Formulations of antibody molecules to influenza virus

ABSTRACT

This disclosure relates to formulations of peptide agents, e.g., antibodies and antigen-binding fragments thereof, that bind hemagglutinin protein of influenza viruses, and methods of their use.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.62/299,162, filed Feb. 24, 2016. The contents of the aforementionedapplication are hereby incorporated by reference in its entirety.

SEQUENCE LISTING

The instant application contains a Sequence Listing which has beensubmitted electronically in ASCII format and is hereby incorporated byreference in its entirety. Said ASCII copy, created on Feb. 20, 2017, isnamed P2029-701110_SL.txt and is 186,341 bytes in size.

BACKGROUND

Influenza is an infectious disease caused by RNA viruses of the familyOrthomyxoviridae (the influenza viruses). Influenza viruses areclassified based on core protein into three genera A, B and C that arefurther divided into subtypes determined by the viral envelopeglycoproteins haemagglutinin (HA) and neuraminidase (NA). Influenza Aviruses infect a range of mammalian and avian species, whereas type Band C infections are largely restricted to humans. Only types A and Bcause human disease of any concern.

High mutation rates and frequent genetic reassortments of the influenzaviruses contribute to great variability of the HA and NA antigens. Minorpoint mutations causing small changes (“antigenic drift”) occurrelatively often. Antigenic drift enables the virus to evade immunerecognition, resulting in repeated influenza outbreaks duringinterpandemic years. Major changes in the HA antigen (“antigenic shift”)are caused by reassortment of genetic material from different influenzaA subtypes. Antigenic shifts resulting in new pandemic strains are rareevents, occurring through reassortment between animal and humansubtypes, for example in co-infected pigs.

Influenza A spreads around the world in seasonal epidemics, resulting inthe deaths of between 250,000 and 500,000 people every year, and up tomillions in some pandemic years. On average 41,400 people died each yearin the United States between 1979 and 2001 from influenza.

SUMMARY

The disclosure is based, at least in part, on the discovery of humananti-HA antibodies comprising functional and structural propertiesdisclosed herein, e.g., antibodies that bind a conserved region orepitope on influenza virus and uses thereof.

Accordingly, the disclosure features formulations comprising bindingagents, e.g., antibody molecules, or preparations, or isolatedpreparations thereof, that bind hemagglutinin (HA) from influenzaviruses. In an embodiment, a binding agent, e.g., an antibody molecule,is broad spectrum, and binds more than one HA, e.g., an HA from one orboth of Group 1 or Group 2 strains of influenza A viruses and/or one ormore strains of influenza B viruses. Therefore, in an embodiment, abinding agent, e.g., an antibody molecule, disclosed herein can treat orprevent infection by a Group 1 influenza virus and a Group 2 influenzavirus. In another embodiment, a binding agent, e.g., an antibodymolecule, disclosed herein can treat or prevent infection by aninfluenza A virus and an influenza B virus. The binding agents, e.g.,antibody molecules, share sufficient structural similarity withantibodies or variable regions disclosed herein, such that they possessfunctional attributes of the antibodies disclosed herein. In anembodiment, the structural similarity can be in terms of a threedimensional structure or a linear amino acid sequence, or both.

In an aspect, the disclosure features a formulation, e.g., apharmaceutical formulation, comprising an anti-HA antibody moleculedescribed herein, e.g., an antibody molecule comprising one, two, orthree heavy chain (HC) CDRs and/or one, two, or three light chain (LC)CDRs of Ab 044, a buffering agent, and a tonicity agent.

In an embodiment, the antibody molecule is present at a concentration ofabout 5 mg/mL to about 150 mg/mL, e.g., about 10 mg/mL to about 100mg/mL, about 15 mg/mL to about 75 mg/mL, about 20 mg/mL to about 60mg/mL, about 20 mg/mL to about 50 mg/mL, about 20 mg/mL to about 30mg/mL, about 15 mg/mL to about 25 mg/mL, about 25 mg/mL to about 35mg/mL, about 25 mg/mL to about 50 mg/mL, about 5 mg/mL to about 20mg/mL, about 8 mg/mL to about 16 mg/mL, about 5 mg/mL to about 50 mg/mL,about 50 mg/mL to about 100 mg/mL, about 40 mg/mL to about 110 mg/mL,about 100 mg/mL to about 150 mg/mL, about 5 mg/mL to about 25 mg/mL,about 10 mg/mL to about 30 mg/mL, about 20 mg/mL to about 40 mg/mL,about 30 mg/mL to about 50 mg/mL, about 40 mg/mL to about 60 mg/mL,about 50 mg/mL to about 70 mg/mL, about 60 mg/mL to about 80 mg/mL,about 70 mg/mL to about 90 mg/mL, about 80 mg/mL to about 100 mg/mL,about 90 mg/mL to about 110 mg/mL, or about 100 mg/mL to about 120mg/mL, e.g., about 150 mg/mL or less, about 100 mg/mL or less, about 50mg/mL or less, about 25 mg/mL or less, about 20 mg/mL or less, about 16mg/mL or less, about 10 mg/mL or less, about 8 mg/mL or less, e.g.,about 5 mg/mL, about 8 mg/mL, about 10 mg/mL, about 15 mg/mL, about 16mg/mL, about 20 mg/mL, about 25 mg/mL, about 30 mg/mL, about 35 mg/mL,about 40 mg/mL, about 45 mg/mL, about 50 mg/mL, about 60 mg/mL, about 70mg/mL, about 80 mg/mL, about 90 mg/mL, about 100 mg/mL, about 110 mg/mL,about 120 mg/mL, about 130 mg/mL, about 140 mg/mL, or about 150 mg/mL.

In an embodiment, the antibody molecule is present at a concentration ofabout 10 to about 40 mg/mL, e.g., about 20 mg/mL to about 30 mg/mL,e.g., about 25 mg/mL. In an embodiment, the antibody molecule is presentat a concentration of about 25 mg/mL. In another embodiment, theantibody molecule is present at a concentration of about 40 mg/mL toabout 60 mg/mL, e.g., about 50 mg/mL. In an embodiment, the antibodymolecule is present at a concentration of about 50 mg/mL. In anotherembodiment, the antibody molecule is present at a concentration of about20 mg/mL to about 60 mg/mL, e.g., about 25 mg/mL to about 50 mg/mL.

In an embodiment, the antibody molecule is present at a concentration ofabout 5 to about 10 mg/mL, e.g., about 8 mg/mL. In an embodiment, theantibody molecule is present at a concentration of about 8 mg/mL. Inanother embodiment, the antibody molecule is present at a concentrationof about 10 mg/mL to about 20 mg/mL, e.g., about 16 mg/mL. In anembodiment, the antibody molecule is present at a concentration of about16 mg/mL. In another embodiment, the antibody molecule is present at aconcentration of about 5 mg/mL to about 20 mg/mL, e.g., about 8 mg/mL toabout 16 mg/mL.

In an embodiment, the antibody molecule is present at a concentration ofabout 25 to about 150 mg/mL, e.g., about 50 mg/mL to about 100 mg/mL,e.g., about 50 mg/mL.

In an embodiment, the antibody molecule comprises a heavy chainimmunoglobulin variable region segment comprising:

-   -   an HC CDR1 comprising the sequence S-Y-A-M-H (SEQ ID NO: 68);    -   an HC CDR2 comprising the sequence        V-V-S-Y-D-G-N-Y-K-Y-Y-A-D-S-V-Q-G (SEQ ID NO: 69); and    -   an HC CDR3 comprising the sequence        D-S-R-L-R-S-L-L-Y-F-E-W-L-S-Q-G-Y-F-N-P (SEQ ID NO: 70).

In an embodiment, the antibody molecule comprises a light chainimmunoglobulin variable region segment comprising:

-   -   an LC CDR1 comprising the sequence Q-S-I-T-F-D-Y-K-N-Y-L-A (SEQ        ID NO: 145);    -   an LC CDR2 comprising the sequence W-G-S-Y-L-E-S (SEQ ID NO:        72); and    -   an LC CDR3 comprising the sequence Q-Q-H-Y-R-T-P-P-S (SEQ ID NO:        73).

In an embodiment, the antibody molecule comprises:

(a) a heavy chain immunoglobulin variable region segment comprising:

-   -   an HC CDR1 comprising the sequence S-Y-A-M-H (SEQ ID NO: 68);    -   an HC CDR2 comprising the sequence        V-V-S-Y-D-G-N-Y-K-Y-Y-A-D-S-V-Q-G (SEQ ID NO: 69); and    -   an HC CDR3 comprising the sequence        D-S-R-L-R-S-L-L-Y-F-E-W-L-S-Q-G-Y-F-N-P (SEQ ID NO: 70); and

(b) a light chain immunoglobulin variable region segment comprising:

-   -   an LC CDR1 comprising the sequence Q-S-I-T-F-D-Y-K-N-Y-L-A (SEQ        ID NO: 145);    -   an LC CDR2 comprising the sequence W-G-S-Y-L-E-S (SEQ ID NO:        72); and    -   an LC CDR3 comprising the sequence Q-Q-H-Y-R-T-P-P-S (SEQ ID NO:        73).

In an embodiment, the antibody molecule comprises a heavy chainimmunoglobulin variable region segment that comprises SEQ ID NO: 25, oran amino acid sequence that differs by no more than 1, 2, 3, 4, or 5amino acids therefrom.

In an embodiment, the antibody molecule comprises a light chainimmunoglobulin variable region segment that comprises SEQ ID NO: 52, oran amino acid sequence that differs by no more than 1, 2, 3, 4, or 5amino acids therefrom.

In an embodiment, the antibody molecule comprises:

(a) a heavy chain immunoglobulin variable region segment that comprisesSEQ ID NO: 25, or an amino acid sequence that differs by no more than 1,2, 3, 4, or 5 amino acids therefrom; and

(b) a light chain immunoglobulin variable region segment that comprisesSEQ ID NO: 52, or an amino acid sequence that differs by no more than 1,2, 3, 4, or 5 amino acids therefrom.

In an embodiment, the antibody molecule comprises:

(a) a heavy chain immunoglobulin variable region segment that comprisesSEQ ID NO: 25; and

(b) a light chain immunoglobulin variable region segment that comprisesSEQ ID NO: 52.

In an embodiment, the buffering agent is present at a concentration ofabout 5 mM to about 150 mM, e.g., about 10 mM to about 100 mM, about 20mM to about 75 mM, about 30 mM to about 50 mM, about 10 mM to about 50mM, about 50 mM to about 100 mM, about 100 mM to about 150 mM, about 10mM to about 30 mM, about 20 mM to about 40 mM, about 30 mM to about 50mM, about 40 mM to about 60 mM, about 50 mM to about 70 mM, about 60 mMto about 80 mM, about 70 mM to about 90 mM, or about 80 mM to about 100mM, e.g., about 150 mM or less, about 100 mM or less, about 75 mM orless, about 50 mM or less, about 25 mM or less, or about 10 mM or less,e.g., about 5 mM, about 10 mM, about 20 mM, about 30 mM, about 40 mM,about 50 mM, about 60 mM, about 70 mM, about 80 mM, about 90 mM, about100 mM, about 110 mM, about 120 mM, about 130 mM, about 140 mM, or about150 mM.

In an embodiment, the buffering agent is present at a concentration ofabout 20 mM to about 60 mM, e.g., about 30 to about 50 mM, e.g., about40 mM. In an embodiment, the buffering agent is present at aconcentration of about 40 mM.

In an embodiment, the buffering agent is a citrate buffer, a phosphatebuffer, or a citrate-phosphate buffer. In an embodiment, the bufferingagent comprises citrate-sodium phosphate. In an embodiment, theformulation comprises citrate-sodium phosphate at a concentration ofabout 20 mM to about 60 mM, e.g., about 30 to about 50 mM, e.g., about40 mM.

In an embodiment, the buffering agent provides a pH of about 5.5 toabout 7, e.g., about 6 to about 6.5, e.g., about 5.5, about 6, about6.5, or about 7. In an embodiment, the buffering agent comprisescitrate-sodium phosphate and provides a pH of about 6 to about 6.5,e.g., about 6 or about 6.5.

In an embodiment, the tonicity agent is present at a concentration ofabout 10 mM to about 500 mM, about 50 mM to about 200 mM, e.g., about 60mM to about 190 mM, about 70 mM to about 180 mM, about 80 mM to about170 mM, about 90 mM to about 160 mM, about 100 mM to about 150 mM, about145 mM to about 155 mM, about 140 mM to about 160 mM, about 135 mM toabout 165 mM, about 130 mM to about 170 mM, about 120 mM to about 180mM, about 110 mM to about 190 mM, about 100 mM to about 200 mM, about 50mM to about 100 mM, about 100 mM to about 150 mM, or about 150 mM toabout 120 mM, e.g., about 200 mM or less, about 150 mM or less, about100 mM or less, or about 75 mM or less, e.g., about 50 mM, about 60 mM,about 70 mM, about 80 mM, about 90 mM, about 100 mM, about 110 mM, about120 mM, about 130 mM, about 140 mM, about 150 mM, about 160 mM, about170 mM, about 180 mM, about 190 mM, or about 200 mM.

In an embodiment, the tonicity agent is present at a concentration ofabout 50 to about 200 mM, about 75 mM to about 150 mM, about 120 mM toabout 180 mM, e.g., about 140 to about 160 mM, e.g., about 150 mM.

In an embodiment, the tonicity agent comprises sodium chloride. In anembodiment, the tonicity agent comprises sodium chloride and is presentat a concentration of about 140 to about 160 mM, e.g., about 150 mM.

In an embodiment, the tonicity agent provides a tonicity (or osmolarity)of about 250 mOsm/L to about 350 mOsm/L, about 260 mOsm/L to about 340mOsm/L, about 270 mOsm/L to about 330 mOsm/L, about 280 mOsm/L to about320 mOsm/L, about 285 mOsm/L to about 310 mOsm/L, or about 290 mOsm/L toabout 300 mOsm/L, e.g., about 250 mOsm/L, about 260 mOsm/L, about 270mOsm/L, about 280 mOsm/L, about 290 mOsm/L, about 300 mOsm/L, about 310mOsm/L, about 320 mOsm/L, about 330 mOsm/L, about 340 mOsm/L, or about350 mOsm/L.

In an embodiment, the formulation has a pH of about 5.5 to about 7,e.g., about 6 to about 6.5, e.g., about 5.5, about 6, about 6.5, orabout 7.

In an embodiment, the formulation comprises:

(a) an antibody molecule described herein at a concentration about 10 toabout 40 mg/mL, e.g., about 20 to about 30 mg/mL, e.g., a concentrationof about 25 mg/mL;

(b) a buffering agent, e.g., citrate-sodium phosphate, at aconcentration about 20 mM to 60 mM, e.g., about 30 to about 50 mM, e.g.,about 40 mM; and

(c) a tonicity agent, e.g., sodium chloride, at a concentration of about75 to about 150 mM, about 120 mM to about 180 mM, e.g., about 140 toabout 160 mM, e.g., a concentration of about 150 mM,

wherein the pH of the formulation is about 5.5 to about 6.5, e.g., about6 or about 6.5.

In an embodiment, the formulation comprises: about 25 mg/mL of anantibody molecule described herein, about 40 mM citrate-sodiumphosphate, about 150 mM sodium chloride, at a pH of about 6.

In an embodiment, the formulation further comprises a surfactant, e.g.,a nonionic surfactant.

In an embodiment, the surfactant is present at a concentration of about0.005% to about 0.1% (w/v), e.g., about 0.01% to about 0.05%, about0.015% to about 0.04%, about 0.02% to about 0.03%, about 0.01% to about0.03%, about 0.02% to about 0.04%, about 0.01% to about 0.025%, about0.025% to about 0.1%, about 0.005% to about 0.05%, or about 0.05% toabout 0.1%, e.g., about 0.1% or less, about 0.075% or less, about 0.05%or less, about 0.025% or less, or about 0.01% or less, e.g., about0.005%, about 0.01%, about 0.015%, about 0.02%, about 0.025%, about0.03%, about 0.035%, about 0.04%, about 0.05%, about 0.06%, about 0.07%,about 0.08%, about 0.09%, or about 0.1%.

In an embodiment, the surfactant is present at a concentration of about0.01% to about 0.05%, e.g., about 0.025%.

In an embodiment, the surfactant is polysorbate 80 (TWEEN® 80). In anembodiment, the surfactant is polysorbate 80 and is present aconcentration of about 0.01% and about 0.05%, e.g., about 0.025%.

In an embodiment, the formulation comprises:

(a) an antibody molecule described herein at a concentration of about 10to about 40 mg/mL, e.g., about 20 to about 30 mg/mL, e.g., about 25mg/mL;

(b) a buffering agent, e.g., citrate-sodium phosphate, at aconcentration of about 20 mM to about 60 mM, e.g., about 30 to about 50mM, e.g., a concentration of about 40 mM;

(c) a tonicity agent, e.g., sodium chloride, at a concentration of about75 mM to about 150 mM, about 120 mM to 180 mM, e.g., about 140 to about160 mM, e.g., a concentration of about 150 mM; and

(d) a surfactant, e.g., polysorbate 80, at a concentration of about0.01% to about 0.04%, e.g., about 0.025%,

wherein the pH of the pharmaceutical composition is about 5.5 to about6.5, e.g., about 6 or about 6.5.

In an embodiment, the formulation comprises about 25 mg/mL of anantibody molecule described herein, about 40 mM citrate-sodiumphosphate, about 150 mM sodium chloride, about 0.025% polysorbate 80, ata pH of about 6.

In an embodiment, the formulation comprises about 25 mg/mL of anantibody molecule described herein, about 40 mM citrate-sodiumphosphate, about 150 mM sodium chloride, about 0.025% polysorbate 80, ata pH of about 6.5.

In an embodiment, the formulation comprises about 25 mg/mL of anantibody molecule described herein, about 40 mM citrate-sodiumphosphate, about 75 mM sodium chloride, about 0.025% polysorbate 80, ata pH of about 6.5.

In an embodiment, the formulation further comprises a stabilizing agent.

In an embodiment, the stabilizing agent is present at a concentration ofabout 0.1% to about 10% (w/v), e.g., about 0.2% to about 5%, about 0.5%to about 1.5%, about 0.5% to about 1%, about 1% to about 2%, e.g., about5% or less, about 4% or less, about 3% or less, about 2% or less, about1% or less, about 0.5% or less, or about 0.2% or less, e.g., about 0.6%,about 0.8%, about 1%, about 1.5%, about 2%, about 3%, about 4%, or about5%.

In an embodiment, the stabilizing agent is an amino acid. In anembodiment, the amino acid is glycine, histidine, arginine, methionine,proline, lysine, glutamic acid, or a combination thereof. In anembodiment, the formulation comprises one, two or all of: glycine,histidine, or arginine. In an embodiment, the amino acid is glycine. Inan embodiment, the formulation comprises glycine, which is present at aconcentration of about 0.5% to about 2%, e.g., about 1%.

In an embodiment, the formulation comprises:

(a) an antibody molecule described herein at a concentration of about 10to about 40 mg/mL, e.g., about 20 to about 30 mg/mL, e.g., about 25mg/mL;

(b) a buffering agent, e.g., citrate-sodium phosphate, at aconcentration of about 20 mM to about 60 mM, e.g., about 30 to about 50mM, e.g., about 40 mM;

(c) a tonicity agent, e.g., sodium chloride, at a concentration of about75 mM to about 150 mM, about 120 mM to about 180 mM, e.g., about 140 toabout 160 mM, e.g., about 150 mM;

(d) a surfactant, e.g., polysorbate 80, at a concentration of about0.01% to about 0.04%, e.g., about 0.025%; and

(c) a stabilizing agent, e.g., glycine, at a concentration of about 0.5%to about 2%, e.g., about 1%,

wherein the pH of the pharmaceutical composition is about 5.5 to about6.5, e.g., about 6 or about 6.5.

In an embodiment, the formulation comprises about 25 mg/mL of anantibody molecule described herein, about 40 mM citrate-sodiumphosphate, about 150 mM sodium chloride, about 0.025% polysorbate 80,about 1% glycine, at a pH of about 6.

In an embodiment, the formulation further comprises a carbohydrate,e.g., a polyol or a sugar. In an embodiment, the carbohydrate issucrose, trehalose, mannitol, dextran, sorbitol, inositol, glucose,fructose, lactose, xylose, mannose, maltose, raffinose, a combinationthereof.

In an embodiment, the formulation further comprises further comprising apolymer, e.g., a hydrophilic polymer. In an embodiment, the polymer is apolyethylene glycol (PEG), dextran, hydroxyl ethyl starch (HETA), orgelatin.

In an embodiment, the formulation further comprises a preservative. Inan embodiment, the preservative is benzyl alcohol, m-cresol, or phenol.

In an embodiment, the level of high molecular weight (HMW) species inthe formulation is less than about 5%, less than about 4%, less thanabout 3%, less than about 2%, or less than 1%, e.g., before storage, orafter storage for at least about 1 week at 4° C., at least about 1 weekat 45° C., at least about 2 weeks at 4° C., at least about 2 weeks at45° C., at least about 3 weeks at 4° C., at least about 3 weeks at 45°C., at least about 4 weeks at 4° C., or at least about 4 weeks at 45° C.In an embodiment, the level of HMW species is less than about 2% beforestorage. In an embodiment, the level of HMW species is less than about2% after storage for 2 weeks at 4° C. In an embodiment, the level of HMWspecies is less than about 5% after storage for 2 weeks at 45° C.

In an embodiment, the level of low molecular weight (LMW) species in theformulation is less than about 5%, less than about 4%, less than about3%, less than about 2%, or less than 1%, e.g., before storage or afterstorage for at least about 1 week at 4° C., at least about 1 week at 45°C., at least about 2 weeks at 4° C., at least about 2 weeks at 45° C.,at least about 3 weeks at 4° C., at least about 3 weeks at 45° C., atleast about 4 weeks at 4° C., or at least about 4 weeks at 45° C. In anembodiment, the level of LMW species is less than about 1% beforestorage. In an embodiment, the level of LMW species is less than about1% after storage for 2 weeks at 4° C. In an embodiment, the level of LMWspecies is less about 2% after storage for 2 weeks at 45° C.

In an embodiment, the level of HMW and LMW species in the formulation isless than about 8%, less than about 7%, less than about 6%, less thanabout 5%, less than about 4%, less than about 3%, less than about 2%, orless than 1%, e.g., before storage, or after storage for at least about1 week at 4° C., at least about 1 week at 45° C., at least about 2 weeksat 4° C., at least about 2 weeks at 45° C., at least about 3 weeks at 4°C., at least about 3 weeks at 45° C., at least about 4 weeks at 4° C.,or at least about 4 weeks at 45° C. In an embodiment, the level of HMWand LMW species is less than about 2% before storage. In an embodiment,the level of HMW and LMW species is less than about 2% after storage for2 weeks at 4° C. In an embodiment, the level of LMW species is less thanabout 6% after storage for 2 weeks at 45° C.

In an embodiment, about 90% or more, about 92% or more, about 94% ormore, about 95% or more, about 96% or more, about 97% or more, about 98%or more, or about 99% or more of the antibody molecules in theformulation are present as monomers, e.g., before storage, or afterstorage for at least about 1 week at 4° C., at least about 1 week at 45°C., at least about 2 weeks at 4° C., at least about 2 weeks at 45° C.,at least about 3 weeks at 4° C., at least about 3 weeks at 45° C., atleast about 4 weeks at 4° C., or at least about 4 weeks at 45° C. In anembodiment, about 98% or more of the antibody molecules in theformulation are present as monomers before storage. In an embodiment,about 98% or more of the antibody molecules in the formulation arepresent as monomers after storage for 2 weeks at 4° C. In an embodiment,about 94% or more of the antibody molecules in the formulation arepresent as monomers after storage for 2 weeks at 45° C.

In an embodiment, the level of monomers, HMW species, or LMW species isdetermined by size exclusion chromatography (SEC), e.g., sizeexclusion-high performance liquid chromatography (SEC-HPLC). In anotherembodiment, the monomeric nature of the antibody molecule is determinedby a binding assay, a surface charge assay, a bioassay, or the ratio ofHMW species to LMW species.

In an embodiment, the purity of the antibody molecule in theformulation, e.g., after storage for two 2 weeks at 4° C., is at leastabout 96%, at least about 97%, at least about 98%, or at least about99%. In an embodiment, the purity of the antibody molecule in theformulation, e.g., after storage for two 2 weeks at 45° C., is at leastabout 90%, at least about 92%, at least about 94%, at least about 96%,at least about 97%, at least about 98%, or at least about 99%. In anembodiment, the purity (or heterogeneity) of the antibody molecule isdetermined by detecting the intact heavy and light chains (e.g., in areduced sample) or intact immunoglobulins (e.g., in a non-reducedsample) in the formulation.

In an embodiment, the purity (or heterogeneity) of the antibody moleculein the formulation is determined by capillary electrophoresis-sodiumdodecyl sulfate (CE-SDS) in a reduced sample. In an embodiment, thepurity (or heterogeneity) of the antibody molecule in the formulation isdetermined by CE-SDS in a non-reduced sample. In an embodiment, thepurity of the antibody molecule in the formulation, e.g., after storagefor two 2 weeks at 4° C., is at least about 98% as determined by CE-SDSin a reduced sample. In an embodiment, the purity of the antibodymolecule in the formulation, e.g., after storage for two 2 weeks at 45°C., is at least about 96% as determined by CE-SDS in a reduced sample.In an embodiment, the purity of the antibody molecule in theformulation, e.g., after storage for two 2 weeks at 4° C., is at leastabout 97% as determined by CE-SDS in a non-reduced sample. In anembodiment, the purity of the antibody molecule in the formulation,e.g., after storage for two 2 weeks at 45° C., is at least about 92% asdetermined by CE-SDS in a non-reduced sample.

In an embodiment, the activity of the antibody molecule is decreased byless than about 25%, less than about 20%, less than about 15%, less thanabout 10%, less than about 5%, or less than about 2%, after storage,e.g., for at least about 1 week, at least about 2 weeks, or at leastabout 3 weeks, e.g., at 4° C. or 45° C. In an embodiment, the activityof the antibody molecule is decreased by less than about 25% afterstorage for about 2 weeks at 45° C. In an embodiment, the activity ofthe antibody molecule is determined by a hemagglutinin (HA) bindingassay, e.g., an HA-binding ELISA.

In an embodiment, the formulation is a liquid formulation (e.g., afrozen or non-frozen liquid formulation). In an embodiment, theformulation is stored as a frozen liquid. In another embodiment, theformulation is a lyophilized formulation.

In an embodiment, the formulation is for use in treating or preventinginfluenza in a subject. In an embodiment, the formulation is for use intreating a subject having influenza. In another embodiment, theformulation is for use in preventing a subject from having influenza. Inan embodiment, the formulation is for intravenous administration.

In another aspect, the disclosure features a device, e.g., an injectiondevice, comprising a formulation described herein, e.g., apharmaceutical formulation described herein.

In yet another aspect, the disclosure features a kit, comprising one ormore containers comprising a formulation described herein, e.g., apharmaceutical formulation described herein, and instructions for use ofthe formulation, e.g., for administration of the formulation to asubject, or for making a solution for administration to a subject.

In another aspect, the disclosure features a container (e.g., a vial oran intravenous (IV) solution bag) comprising an anti-HA antibodymolecule described herein or a formulation (e.g., a pharmaceuticalformulation described herein) comprising an anti-HA antibody moleculedescribed herein.

In an embodiment, the container is a vial, e.g., a glass vial. In anembodiment, the container (e.g., vial) comprises about 10 mg/mL to about100 mg/mL, e.g., about 20 mg/mL to about 60 mg/mL (e.g., about 25 mg/mLto about 50 mg/mL) of the antibody molecule. In an embodiment, thecontainer (e.g., vial) comprises about 10 mL to about 60 mL, e.g., about20 mL to about 40 mL, of the formulation. In an embodiment, thecontainer (e.g., vial) is a first (or primary) container, e.g., forstoring the antibody molecule or formulation.

The antibody molecule or formulation can be transferred into a second(or secondary) container before use. In an embodiment, the second (orsecondary) container is suitable, or includes a solution that issuitable, for administration, e.g., intravenous administration. In anembodiment, the second (or secondary) container includes a solutionsuitable for intravenous administration. In an embodiment, the solutioncomprises saline, optionally, further comprises dextrose. In anembodiment, the solution (e.g., saline) does not comprise dextrose. Forexample, an amount equal to one dose of the antibody molecule can betransferred into a container suitable for IV administration. In anembodiment, 1 to 10 vials (e.g., 1 to 8 vials, 1 to 6 vials, 1 to 4vials, 1 to 2 vials, 6 to 8 vials, 4 to 8 vials, or 2 to 8 vials) of theformulation are transferred (e.g., diluted) into an IV solution bag,e.g., containing saline with or without dextrose.

In an embodiment, the container is a container suitable for IVadministration (e.g., an IV solution bag). In an embodiment, the amountof the antibody molecules in the container (e.g., IV solution bag)equals to the amount of the antibody molecules in 1 to 10 vials (e.g., 1to 8 vials, 1 to 6 vials, 1 to 4 vials, 1 to 2 vials, 6 to 8 vials, 4 to8 vials, or 2 to 8 vials) of the formulation as described above. In anembodiment, the container (e.g., IV solution bag) comprises about 2000mg to about 5000 mg, e.g., about 2300 mg to about 4600 mg, of theantibody molecule, e.g., in a solution suitable for IV administration(e.g., saline with or without dextrose).

In an embodiment, the container suitable for IV administration (e.g., IVsolution bag) is not a second (or secondary) container (e.g., is a first(or primary) container, e.g., where the antibody molecule is stored),and comprises about 5 mg/mL to about 25 mg/mL, e.g., about 8 mg/mL toabout 16 mg/mL of the antibody molecule. In an embodiment, the container(e.g., IV solution bag) comprises about 100 mL to about 400 mL (e.g.,about 200 mL to about 300 mL) of a solution (e.g., a solution suitablefor IV administration) comprising the antibody molecule. In anembodiment, the container (e.g., IV solution bag) comprises about 2000mg to about 5000 mg, e.g., about 2000 mg to about 4000 mg or about 2300mg to about 4600 mg, of the antibody molecule.

In another aspect, the disclosure features a method of preparing acomposition (e.g., a solution) for administration to a subject. Themethod comprises combining a formulation described herein with asolution suitable for intravenous administration.

In an embodiment, the solution comprises saline, optionally, furthercomprises dextrose. In an embodiment, the solution does not comprisedextrose. In an embodiment, about 2000 mg to about 5000 mg of theantibody molecule is combined with the solution. In another embodiment,about 2300 mg to about 4600 mg of the antibody molecule is combined withthe solution. In yet another embodiment, about 2000 mg to about 4000 mgof the antibody molecule is combined with the solution. In anembodiment, the formulation is combined with the solution in anintravenous (IV) solution bag.

In still another aspect, the disclosure features a method of treating orpreventing influenza, the method comprising administering to a subjecthaving influenza, or at risk of having influenza, an effective amount ofa formulation described herein, e.g., a pharmaceutical formulationdescribed herein, thereby treating or preventing influenza.

In an aspect, the disclosure features a formulation described herein,e.g., a pharmaceutical formulation described herein, for use in treatingor preventing influenza in a subject.

Anti-HA Antibody Molecules

Various anti-HA antibody molecules, or preparations, or isolatedpreparations thereof, can be included in a formulation (e.g.,pharmaceutical formulation) described herein.

In an embodiment, the antibody molecule comprises one or more (e.g., 2,3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or all) of thefollowing properties:

(a) it fails to produce any escape mutants as determined by the failureof a viral titer to recover following at least 10, 9, 8, 7, 6, or 5rounds of serial infections in cell culture with a mixture of theantibody molecule and an influenza A virus, e.g., a Group 1 strain,e.g., an H1N1 strain, e.g., A/South Carolina/1/1918, A/PuertoRico/08/1934, or A/California/04/2009, an H5N1 strain, e.g.,A/Indonesia/5/2005 or A/Vietnam/1203/2004, or an H7N9, e.g.,A/Anhui/1/2013 or A/Shanghai/1/2013;

(b) it produces fewer escape mutants than does a reference anti-HAantibody molecule, e.g., Ab 67-11, FI6, FI28, C179, F10, CR9114, orCR6261, e.g., when tested by the method described in (a);

(c) it binds with high affinity to an HA of at least 1, 2, 3, 4, or 5influenza subtypes of Group 1 and at least 1, 2, 3, 4, or 5 influenzasubtypes of Group 2;

(d) it prevents infection by at least 1, 2, 3, 4 or 5 influenza subtypesof Group 1, and by at least 1, 2, 3, 4 or 5 influenza subtypes of Group2;

(e) it inhibits fusogenic activity of the targeted HA;

(f) it treats or prevents infection by a Group 1 virus, such as wherethe virus is an H1, H5, or H9 virus; and it treats or prevents infectionby a Group 2 virus, such as where the virus is an H3 or H7 virus;

(g) it treats or prevents infection by an influenza A H1N1 strain, aninfluenza A H3N2 strain, or both;

(h) it is effective for prevention or treatment of infection, e.g., inhumans or mice, with H1N1 or H3N2 when administered at 50 mg/kg, 25mg/kg, 10 mg/kg, 6 mg/kg, 5 mg/kg, 4 mg/kg, 3 mg/kg, 2 mg/kg, or 1mg/kg;

(i) it treats or prevents infection by an influenza A H5N1 strain, aninfluenza A H7N9 strain, or both;

(j) it is effective for prevention or treatment of infection, e.g., inhumans or mice, with H5N1 or H7N9 when administered at 50 mg/kg, 25mg/kg, 10 mg/kg, 6 mg/kg, 5 mg/kg, 4 mg/kg, 3 mg/kg, 2 mg/kg, or 1mg/kg;

(k) the concentration of antibody molecule required for 50%neutralization of influenza A virus is less than 10 μg/mL;

(l) it treats or prevents infection by an influenza B virus, e.g.,B/Wisconsin/1/2010;

(m) it is effective for prevention or treatment of infection, e.g., inhumans or mice, with an influenza B virus, e.g., B/Wisconsin/1/2010,when administered at 10 mg/kg, 6 mg/kg, 4 mg/kg, 3 mg/kg, 2 mg/kg, or 1mg/kg;

(n) the concentration of antibody molecule required for 50%neutralization of influenza B virus, e.g., B/Wisconsin/1/2010, virus isless than 10 μg/mL;

(o) it prevents or minimizes a secondary infection (e.g., secondarybacterial infection) or an effects thereof on a subject;

(p) it is effective for preventing or minimizing secondary infection(e.g., secondary bacterial infection) or effects thereof on a subjectwhen administered at 50 mg/kg, 25 mg/kg, 10 mg/kg, 6 mg/kg, 5 mg/kg, 4mg/kg, 3 mg/kg, 2 mg/kg, or 1 mg/kg;

(q) it binds an epitope which comprises or consists of the hemagglutinintrimer interface; and

(r) it binds an epitope other than that bound by a reference anti-HAantibody molecule, e.g., Ab 67-11, FI6, FI28, C179, F10, CR9114, orCR6261, e.g., as determined by a method described herein, e.g., astructural analysis (e.g., by X-ray crystallography or NMR spectroscopy)or a competition assay (e.g., by ELISA); or

(s) it binds to an epitope, e.g., it has an epitope that overlaps withor is the same as, of an antibody disclosed herein, e.g., as determinedby a method described herein (e.g., a mutational analysis or a crystalstructure analysis).

In an embodiment, the antibody molecule has one, two, or all of thefollowing characteristics: (i) the antibody molecule prevents infectionby at least 1, 2, 3, 4 or 5 influenza subtypes of Group 1, and by atleast 1, 2, 3, 4 or 5 influenza subtypes of Group 2; (ii) theconcentration of the antibody molecule required for 50% neutralizationof influenza A virus is less than 10 μg/mL; or (iii) the antibodymolecule binds an epitope that comprises or consists of thehemagglutinin trimer interface.

In an embodiment, the antibody molecule treats or prevents infection bya Group 1 virus, such as where the virus is an H1, H2, H5, H6, H8, H9,H12, H11, H13, H16, or H17 virus; and treats or prevents infection by aGroup 2 virus, such as where the virus is an H3, H4, H7, H10 or H15virus. In an embodiment, the antibody molecule prevents infection by atleast 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11 influenza subtypes of Group 1,and by at least 1, 2, 3, 4, 5 or 6 influenza subtypes of Group 2. In anembodiment, the antibody molecule treats or prevents infection by one ormore of H1N1, H2N2, H5N1, or H9N2, and also treats or prevents infectionby one or more of H3N2, H7N7, or H7N9.

In an embodiment, the antibody molecule binds, and in an embodiment,neutralizes: at least one strain from the Group 1 H1, e.g., H1a or H1b,cluster and at least one strain from the Group 2 H3 or H7 cluster. In anembodiment, the antibody molecule, binds, and in an embodiment,neutralizes: at least one strain from the Group 1 H1, e.g., H1a or H1b,cluster and at least one influenza B strain, e.g., B/Wisconsin/1/2010.In an embodiment, the antibody molecule binds, and in an embodiment,neutralizes: at least one strain from the Group 2 H3 or H7 cluster andat least one influenza B strain, e.g., B/Wisconsin/1/2010. In anembodiment, the antibody molecule binds, and in an embodiment,neutralizes: at least one strain from the Group 1 H1, e.g., H1a or H1b,cluster, at least one strain from the Group 2 H3 or H7 cluster, and atleast one influenza B strain, e.g., B/Wisconsin/1/2010. In anembodiment, the antibody molecule treats or prevents infection by one ormore of influenza B viruses, e.g., B/Wisconsin/1/2010.

In an embodiment, the antibody molecule is not an anti-HA antibodymolecule previously described in the art. For example, the antibodymolecule is other than one or more or all of Ab 67-11 (U.S. ProvisionalApplication No. 61/645,453, U.S. Application Publication No.2013/0302348, and International Application Publication No. WO2013/169377), FI6 (FI6, as used herein, refers to any specificallydisclosed FI6 sequence in U.S. Application Publication Nos. 2010/0080813or 2011/0274702, International Application Publication No.WO2013/011347, or Corti et al., Science 333:850-856, 2011; FIG. 4), FI28(U.S. Application Publication No. 2010/0080813), C179 (Okuno et al., J.Virol. 67:2552-1558, 1993), F10 (Sui et al., Nat. Struct. Mol. Biol.16:265, 2009), CR9114 (Dreyfus et al., Science 337(6100):1343-1348,2012), or CR6261 (Ekiert et al., Science 324:246-251, 2009).

In an embodiment, the antibody molecule neutralizes infection with H1N1and H3N2 in vitro. In another embodiment, the antibody moleculeneutralizes infection with H1N1 and H3N2 in vivo. In an embodiment, theantibody molecule neutralizes infection with H5N1 in vitro. In anotherembodiment, the antibody molecule neutralizes infection with H5N1 invivo. In an embodiment, the antibody molecule neutralizes infection withH7N9 in vitro. In another embodiment, the antibody molecule neutralizesinfection with H7N9 in vivo. In an embodiment, the antibody moleculeneutralizes infection with an influenza B virus, e.g.,B/Wisconsin/1/2010, in vitro. In another embodiment, the antibodymolecule neutralizes infection with an influenza B virus, e.g.,B/Wisconsin/1/2010, in vivo.

In another embodiment, the concentration of the antibody moleculerequired for 50% neutralization of influenza A virus is 10 μg/mL orless, such as 9 μg/mL or less, 8 μg/mL or less, 7 μg/mL or less, 6 μg/mLor less, or 5 μg/mL or less. In another embodiment, the concentration ofthe antibody molecule required for 60% neutralization of influenza Avirus, 50% neutralization of influenza A virus, or 40% neutralization ofinfluenza A virus is 10 μg/mL or less, such as 9 μg/mL or less, 8 μg/mLor less, 7 μg/mL or less, 6 μg/mL or less, or 5 μg/mL or less.

In yet another embodiment, the antibody molecule is effective forprevention or treatment of infection, e.g., in humans or mice, with H1N1or H3N2, such as when administered at 50 mg/kg, 25 mg/kg, 10 mg/kg, 6.0mg/kg, 5.0 mg/kg, 4.0 mg/kg, 3.0 mg/kg, 2.0 mg/kg, 1.0 mg/kg or less. Instill another embodiment, the antibody molecule is effective forprevention or treatment of infection, e.g., in humans or mice, with H5N1or H7N9, such as when administered at 50 mg/kg, 25 mg/kg, 10 mg/kg, 6.0mg/kg, 5.0 mg/kg, 4.0 mg/kg, 3.0 mg/kg, 2.0 mg/kg, 1.0 mg/kg or less. Inanother embodiment, the antibody molecule is effective for the treatmentor prevention of a Group 1 virus, where the Group 1 virus is H1, H5, orH9, and in another embodiment, the anti-HA antibody molecule, iseffective for the treatment or prevention of a Group 2 virus, where theGroup 2 virus is H3 or H7.

In another embodiment, the concentration of the antibody moleculerequired for 50% neutralization of influenza B virus, e.g.,B/Wisconsin/1/2010, is 10 μg/mL or less, such as 9 μg/mL or less, 8μg/mL or less, 7 μg/mL or less, 6 μg/mL or less, or 5 μg/mL or less. Inanother embodiment, the concentration of the antibody molecule requiredfor 60% neutralization of influenza B virus, e.g., B/Wisconsin/1/2010,50% neutralization of influenza B virus, e.g., B/Wisconsin/1/2010, or40% neutralization of influenza B virus, e.g., B/Wisconsin/1/2010, is 10μg/mL or less, such as 9 μg/mL or less, 8 μg/mL or less, 7 μg/mL orless, 6 μg/mL or less, or 5 μg/mL or less.

In an embodiment, the antibody molecule comprises one or both of thefollowing properties: (i) it fails to produce any escape mutants asdetermined by the failure of a viral titer to recover following at least10, 9, 8, 7, 6, or 5 rounds of serial infections in cell culture with amixture of the antibody molecule and an influenza virus (e.g., aninfluenza A virus, e.g., a Group 1 strain, e.g., an H1N1 strain, e.g.,A/South Carolina/1/1918, A/Puerto Rico/08/1934, or A/California/04/2009,or an H5N1 strain, e.g., A/Indonesia/5/2005 or A/Vietnam/1203/2004, oran influenza B virus, e.g., B/Wisconsin/1/2010); or (ii) it producesfewer escape mutants than does a reference anti-HA antibody molecule,such as Ab 67-11, FI6, FI28, C179, F10, CR9114, or CR6261, such as whentested by the method described in (i).

In another embodiment, the antibody molecule is a full length tetramericantibody, a single chain antibody (scFv), a F(ab′)₂ fragment, a Fabfragment, or an Fd fragment. In another embodiment, the heavy chain ofthe antibody molecule is a γ1 heavy chain, and in yet anotherembodiment, the light chain of the antibody molecule is a κ light chainor a λ light chain. In yet another embodiment, the antibody molecule isan IgG1 antibody.

In an embodiment, the antibody molecule binds an epitope that has one,two, three, four, five, or all of, the following properties a)-f): a) itincludes one, two, or all of, H3 HA1 residues N38, 1278, and D291; b) itincludes H3 HA2 residue N12; c) it does not include one, two or all of,H3 HA1 residues Q327, T328, and R329; d) it does not include one, two,three, four, or all of, H3 HA2 residues G1, L2, F3, G4, and D46; e) itincludes one, two, or all of, H3 HA1 residues T318, R321, and V323; orf) it includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,17, or all of, H3 HA2 residues A7, E11, I18, D19, G20, W21, L38, K39,T41, Q42, A43, I45, I48, N49, L52, N53, I56, and E57.

In an embodiment the antibody molecule has properties: a) and b). In anembodiment the antibody molecule has properties: c) and d). In anembodiment the antibody molecule has properties: a); and c) or d). In anembodiment the antibody molecule has properties: b); and c) or d). In anembodiment the antibody molecule has properties: c); and a) or b). In anembodiment the antibody molecule has properties: d); and a) or b). In anembodiment the antibody molecule has properties: a), b), c) and d). Inan embodiment the antibody molecule has properties: a), b), c), d), e),and f).

In an embodiment, the antibody molecule has a K_(D) for H3 of equal toor less than 10⁻⁶ nM, wherein said K_(D) is increased by at least 2, 5,10, or 100 fold, by a mutation or mutations in any of: a) H3 HA1residues N38, 1278, or D291; b) H3 HA2 residue N12; c) H3 HA1 residuesT318, R321, or V323; or d) H3 HA2 residues A7, E11, I18, D19, G20, W21,L38, K39, T41, Q42, A43, I45, I48, N49, L52, N53, 156, or E57. In anembodiment, the antibody molecule has a K_(D) for H3 of equal to or lessthan 10⁻⁶ nM, wherein said K_(D) is increased by no more than 2, or 5fold, by a mutation or mutations in any of: e) H3 HA1 residues Q327,T328, or R329; or f) H3 HA2 residues G1, L2, F3, G4, or D46.

In an embodiment, the antibody molecule binds an epitope that has one,two, three, four, five, or all of, the following properties aa)-ff): aa)it includes one, two, or all of, H1 HA1 residues H31, N279, and 5292;bb) it includes H1 HA2 residue G12; cc) it does not include one or bothof H1 HA1 residues Q328 and 5329; dd) it does not include one, two,three, four, or all of, H1 HA2 residues G1, L2, F3, G4, and D46; ee) itincludes one, two, or all of, H1 HA1 residues T319, R322, and 1324 arebound by both Ab 044 and FI6; or ff) it includes 1, 2, 3, 4, 5, 6, 7, 8,9, 10, 11, 12, 13, 14, 15, 16, 17, or all of, H1 HA2 residues A7, E11,I18, D19, G20, W21, Q38, K39, T41, Q42, N43, 145, 148, T49, V52, N53,156, and E57. In an embodiment, the antibody molecule has properties:aa) and bb). In an embodiment, the antibody molecule has properties: cc)and dd). In an embodiment, the antibody molecule has properties: aa);and cc) or dd). In an embodiment, the antibody molecule has properties:bb); and cc) or dd). In an embodiment, the antibody molecule hasproperties: cc); and aa) or bb). In an embodiment, the antibody moleculehas properties: dd); and aa) or bb). In an embodiment, the antibodymolecule has properties: aa), bb), cc) and dd). In an embodiment, theantibody molecule has properties: aa), bb), cc), dd), ee), and ff).

In an embodiment, the antibody molecule has a K_(D) for H1 of equal toor less than 10⁻⁶ nM, wherein said K_(D) is increased by at least 2, 5,10, or 100 fold, by a mutation or mutations in any of: aa) H1 HA1residues H31, N279, and 5292; bb) H1 HA2 residue G12; cc) H1 HA1residues T319, R322, and 1324; or dd) H1 HA2 residues A7, E11, I18, D19,G20, W21, Q38, K39, T41, Q42, N43, 145, 148, T49, V52, N53, 156, andE57. In an embodiment, the antibody molecule has a K_(D) for H1 of equalto or less than 10⁻⁶ nM, wherein said K_(D) is increased by no more than2, or 5 fold, by a mutation or mutations in any of: ee) H1 HA1 residuesQ328 and S329; or ff) H1 HA2 residues G1, L2, F3, G4, and D46.

In an embodiment, the antibody molecule has one, two, three or all ofthe following properties: a) and aa); b) and bb); c) and cc); or d) anddd), as described above. In an embodiment, the antibody molecule hasproperties c), cc), d), and dd), as described above.

In an embodiment, the antibody molecule comprises one or both of: aheavy chain variable region comprising at least, or more than, 60, 65,70, 75, 80, 85, 87, 90, 95, 98 or 99 percent homology with a heavy chainvariable region from Table 3, Table 4A, Table 4B, FIG. 2, FIG. 5 or FIG.7; and a light chain variable region comprising at least, or more than,60, 65, 70, 75, 80, 85, 87, 90, 95, 98 or 99 percent homology with lightchain variable region from Table 3, Table 4A, Table 4B, FIGS. 3A-3B,FIGS. 6A-6B or FIG. 7.

In an embodiment, the antibody molecule comprises a heavy chain variableregion 25 (SEQ ID NO: 25), or a structurally or functionally relatedvariable heavy chain region as described herein. In an embodiment, theantibody molecule comprises a light chain variable region 52 (SEQ ID NO:52), 155 (SEQ ID NO: 155), or 45 (SEQ ID NO: 45), or a structurally orfunctionally related variable light chain region as described herein. Inan embodiment, the antibody molecule comprises a heavy chain variableregion 25 (SEQ ID NO: 25), or a structurally or functionally relatedvariable heavy chain region as described herein; and a light chainvariable region 52 (SEQ ID NO: 52), 155 (SEQ ID NO: 155), or 45 (SEQ IDNO: 45), or a structurally or functionally related variable light chainregion as described herein.

In an embodiment, the antibody molecule comprises a heavy chain variableregion comprising one, two, or all of CDR1, CDR2, and CDR3, from heavychain variable region 25 (SEQ ID NO: 25), or a structurally orfunctionally related variable heavy chain region as described herein. Inan embodiment, the antibody molecule comprises a light chain variableregion comprising one, two, or all of CDR1, CDR2, and CDR3, from lightchain variable region 52 (SEQ ID NO: 52), 155 (SEQ ID NO:155), or 45(SEQ ID NO:45), or a structurally or functionally related sequence asdescribed herein. In an embodiment, the antibody molecule comprises aheavy chain variable region comprising one, two, or all of CDR1, CDR2,and CDR3, from heavy chain variable region 25 (SEQ ID NO: 25), or astructurally or functionally related variable heavy chain region asdescribed herein; and a light chain variable region comprising one, two,or all of CDR1, CDR2, and CDR3, from light chain variable region 52 (SEQID NO: 52), 155 (SEQ ID NO: 155), or 45 (SEQ ID NO: 45), or astructurally or functionally related variable light chain region asdescribed herein.

In an embodiment, the antibody molecule comprises a heavy chain variableregion from FIG. 2 or FIG. 5 or a structurally or functionally relatedvariable heavy chain region as described herein. In an embodiment, theantibody molecule comprises a light chain variable region from FIGS.3A-3B or FIGS. 6A-6B or a structurally or functionally related variablelight chain region as described herein.

In an embodiment, the antibody molecule comprises one, two, or all of, aCDR1, CDR2, and CDR3 from a heavy chain variable region from FIG. 2 orFIG. 5, or a structurally or functionally related sequences as describedherein. In an embodiment, the antibody molecule comprises one, two, orall of, a CDR1, CDR2, and CDR3 from a light chain variable region fromFIGS. 3A-3B or FIGS. 6A-6B, or a structurally or functionally relatedsequences as described herein.

In an embodiment the antibody molecule comprises one, two or all of, HCCDR1, HC CDR2, and HC CDR3 and one, two or all of, LC CDR1, LC CDR2, andLC CDR3 from an antibody disclosed in Table 3, or a structurally orfunctionally related sequence as described herein.

In another embodiment, the antibody molecule comprises the light chainLC45 (SEQ ID NO: 45). In yet another embodiment, the antibody comprisesthe light chain LC45, and the heavy chain HC25 (SEQ ID NO: 25) or HC24(SEQ ID NO: 24). In an embodiment, the antibody molecule comprises thelight chain LC45 (SEQ ID NO: 45) and the heavy chain HC25 (SEQ ID NO:25). In yet another embodiment, the antibody molecule comprises lightchain LC52 (SEQ ID NO: 52) and heavy chain HC25 (SEQ ID NO: 25).

In an embodiment the antibody molecule comprises one or both of: a) oneor more framework regions (FRs) from heavy chain disclosed herein, e.g.,one or more or all of FR1, FR2, FR3, or FR4, or FR sequences that differindividually, or collectively, by no more than 1, 2, 3, 4, of 5 aminoacid residues, e.g., conservative residues, from a heavy chain disclosedherein; and b) one or more framework regions (FRs) from light chaindisclosed herein, e.g., one or more or all of FR1, FR2, FR3, or FR4, orFR sequences that differ individually, or collectively, by no more than1, 2, 3, 4, of 5 amino acid residues, e.g., conservative residues, froma light chain disclosed herein.

In an embodiment, the antibody molecule comprises: (a) a heavy chainimmunoglobulin variable domain comprising a sequence at least 60, 70,80, 85, 87, 90, 95, 97, 98, or 99, e.g., 90%, homologous, to a heavychain consensus sequence provided herein, e.g., the heavy chainconsensus sequence provided in FIG. 2 or FIG. 5, e.g., the heavy chainconsensus sequence provided in FIG. 2, SEQ ID NO: 161; and (b) a lightchain immunoglobulin variable domain comprising a sequence at least 60,70, 80, 85, 87, 90, 95, 97, 98, or 99, e.g., 95%, homologous, to a lightchain consensus sequence provided herein, e.g., the light chainconsensus sequence provided in FIGS. 3A-3B or FIG. 6, e.g., the lightchain consensus sequence provided in FIGS. 3A-3B, SEQ ID NO: 62.

For example, in an embodiment, the antibody molecule disclosed hereincomprises one or both of: (a) a heavy chain immunoglobulin variabledomain comprising the sequence of SEQ ID NO: 161, or a sequence at least87% identical to SEQ ID NO: 161; and (b) a light chain immunoglobulinvariable domain comprising the sequence SEQ ID NO: 62, or a sequence atleast 95% identical to SEQ ID NO: 62.

In another embodiment, the antibody molecule comprises: (a) a heavychain immunoglobulin variable domain comprising the sequence of SEQ IDNO:161, or a sequence at least 87% identical to SEQ ID NO: 161; and (b)a light chain immunoglobulin variable domain comprising the sequence SEQID NO: 62, or a sequence at least 95% identical to SEQ ID NO: 62,wherein said antibody molecule: (i) fails to produce any escape mutantsas determined by the failure of a viral titer to recover following atleast 10, 9, 8, 7, 6, or 5 rounds of serial infections in cell culturewith a mixture of the antibody molecule and an influenza virus (e.g., aninfluenza A virus, e.g., a Group 1 strain, e.g., an H1N1 strain, e.g.,A/South Carolina/1/1918, A/Puerto Rico/08/1934, or A/California/04/2009,or an H5N1 strain, e.g., A/Indonesia/5/2005 or A/Vietnam/1203/2004, oran influenza B virus, e.g., B/Wisconsin/1/2010); and (ii) produces fewerescape mutants than does a reference anti-HA antibody molecule, e.g., Ab67-11, FI6, FI28, C179, F10, CR9114, or CR6261, such as when tested bythe method described in (i).

In an embodiment, the antibody molecule comprises one or both of: (a) aheavy chain immunoglobulin variable region comprising the sequence ofSEQ ID NO: 161, or a sequence that differs from SEQ ID NO: 161 by notmore than 1, 2, 3, 4, 5, 6, 8, 10, 11, 12, 13, 14, 15 or 16, e.g., by nomore than 2, 3, 4, or 5 amino acids, e.g., conservative amino acids; and(b) a light chain immunoglobulin variable domain comprising the sequenceSEQ ID NO: 62, or a sequence that differs from SEQ ID NO: 62 thatdiffers by no more than 1, 2, 3, 4 or 5 amino acids, e.g., conservativeamino acids.

In an embodiment, the 1, 2, 3, 4, 5, 6, 8, 10, 11, 12, 13, 14, 15 or 16amino acid differences, e.g., conservative amino acid differences, inthe heavy chain immunoglobulin variable region are in the FR regions ofthe heavy chain immunoglobulin variable domain. In another embodiment,the 1, 2, 3, 4 or 5 amino acid differences, e.g., conservative aminoacid differences, in the light chain immunoglobulin variable domain arein the FR regions of the light chain immunoglobulin variable domain. Inan embodiment, the amino acid differences in the heavy chainimmunoglobulin variable region, or in the light chain immunoglobulinvariable region, are conservative amino acid changes.

In an embodiment, the antibody molecule comprises one or both of: a) oneor more framework regions (FRs) from heavy chain consensus sequencedisclosed herein, e.g., one or more or all of FR1, FR2, FR3, or FR4, orsequences that differ individually, or collectively, by no more than 1,2, 3, 4, of 5 amino acid residues, e.g., conservative residues, from aheavy chain variable region consensus sequence disclosed herein; and b)one or more framework regions (FRs) from light chain consensus sequencedisclosed herein, e.g., one or more or all of FR1, FR2, FR3, or FR4, orsequences that differ individually, or collectively, by no more than 1,2, 3, 4, of 5 amino acid residues, e.g., conservative residues, from alight chain variable region consensus disclosed herein.

In an embodiment, the antibody molecule binds to an epitope, e.g., anepitope that overlaps with or is the same as, of an antibody disclosedherein, e.g., as determined by mutational analysis or crystal structureanalysis.

In an embodiment, the antibody molecule competes with a referenceantibody molecule, e.g., an antibody molecule described herein, forbinding to a substrate, e.g., an HA.

The HA can be from a Group 1 strain, e.g., HA1 or HA5, e.g., from anH1N1 strain, e.g., A/South Carolina/1/1918, A/Puerto Rico/08/1934, orA/California/04/2009, or an H5N1 strain, e.g., A/Indonesia/5/2005 orA/Vietnam/1203/2004 Binding to the same epitope, or a portion thereof,can be shown by one or more of: a) mutational analysis, e.g., binding toHA, or binding affinity for HA, is decreased or abolished if a residueis mutated; b) analysis, e.g., comparison, of the crystal structure ofthe antibody molecule and HA and the crystal structure of a referenceantibody and HA, e.g., to determine the touch points of each; c)competition of the two antibodies for binding to HA, e.g., HA1 or HA5,from, e.g., an H1N1 strain, e.g., A/South Carolina/1/1918, A/PuertoRico/08/1934, or A/California/04/2009, or an H5N1 strain, e.g.,A/Indonesia/5/2005 or A/Vietnam/1203/2004; or d) (c) and one or both of(a) and (b).

Competition between the antibody molecule and a reference antibodymolecule can be determined by evaluating the ability of the antibodymolecule or the reference antibody molecule to decrease binding of theother to a substrate, e.g., HA, e.g., HA1 or HA5, from, e.g., an H1N1strain, e.g., A/South Carolina/1/1918, A/Puerto Rico/08/1934, orA/California/04/2009, or an H5N1 strain, e.g., A/Indonesia/5/2005 orA/Vietnam/1203/2004. Reduction of the ability to bind can be evaluatedby methods in the art. Reduction of the ability to bind can beevaluated, e.g., by one or more of: a) BIAcore analysis; b) ELISA assay;or c) flow cytometry. The antibody molecule can compete with thereference antibody such that binding of the reference antibody isdecreased by 50% or more.

In an embodiment, the antibody molecule binds to the same epitope, or aportion thereof, which the reference antibody molecule binds. In anembodiment, the antibody molecule does not bind to the same epitope, ora portion thereof, which the reference antibody molecule binds.

In an embodiment, the antibody molecule comprises a structural orfunctional property of Ab 044.

In an embodiment, the antibody molecule competes with a referenceantibody molecule, e.g., an antibody molecule described herein, forbinding to a substrate, e.g., an HA. The reference antibody molecule canbe: a) an antibody molecule comprising: i) a heavy chain immunoglobulinvariable region segment comprising a CDR1 comprising the sequenceS-Y-A-M-H (SEQ ID NO: 68); a CDR2 comprising the sequenceV-V-S-Y-D-G-N-Y-K-Y-Y-A-D-S-V-Q-G (SEQ ID NO: 69); and a CDR3 comprisingthe sequence D-S-R-L-R-S-L-L-Y-F-E-W-L-S-Q-G-Y-F-N-P (SEQ ID NO:70); andii) a light chain variable region segment comprising: a CDR1 comprisingthe sequence Q-S-I-T-F-D-Y-K-N-Y-L-A (SEQ ID NO:145); a CDR2 comprisingthe sequence W-G-S-Y-L-E-S (SEQ ID NO:72); and a CDR3 comprising thesequence Q-Q-H-Y-R-T-P-P-S (SEQ ID NO:73); b) an antibody moleculecomprises one or both of: (i) a heavy chain immunoglobulin variableregion segment comprising SEQ ID NO: 25; and (ii) a light chain variableregion segment comprising SEQ ID NO: 52; or c) Ab 044.

In an embodiment, the antibody molecule binds to the same epitope, or aportion thereof, on HA, as does a reference antibody molecule, e.g., anantibody molecule disclosed herein. The reference antibody molecule canbe: a) an antibody molecule comprising: i) a heavy chain immunoglobulinvariable region segment comprising a CDR1 comprising the sequenceS-Y-A-M-H (SEQ ID NO: 68); a CDR2 comprising the sequenceV-V-S-Y-D-G-N-Y-K-Y-Y-A-D-S-V-Q-G (SEQ ID NO: 69); and a CDR3 comprisingthe sequence D-S-R-L-R-S-L-L-Y-F-E-W-L-S-Q-G-Y-F-N-P (SEQ ID NO: 70);and ii) a light chain variable region segment comprising: a CDR1comprising the sequence Q-S-I-T-F-D-Y-K-N-Y-L-A (SEQ ID NO: 145); a CDR2comprising the sequence W-G-S-Y-L-E-S (SEQ ID NO: 72); and a CDR3comprising the sequence Q-Q-H-Y-R-T-P-P-S (SEQ ID NO: 73); b) anantibody molecule comprises one or both of: (i) a heavy chainimmunoglobulin variable region segment comprising SEQ ID NO: 25; and(ii) a light chain variable region segment comprising SEQ ID NO:52; orc) Ab 044.

In an embodiment, the an antibody molecule comprises one or both of: aheavy chain variable region comprising at least 60, 70, 80, 85, 90, 95,98 or 99 percent homology with SEQ ID NO: 25; and a light chain variableregion comprising at least 60, 70, 80, 85, 90, 95, 98 or 99 percenthomology with SEQ ID NO: 52.

In an embodiment, the antibody molecule, comprises one or both of: aheavy chain variable region comprising at least 60, 70, 80, 85, 90, 95,98 or 99 percent homology with SEQ ID NO: 25; and a light chain variableregion comprising at least 60, 70, 80, 85, 90, 95, 98 or 99 percenthomology with SEQ ID NO: 52, wherein, each HC CDR differs by no morethan 1, 2, 3, 4 or 5 amino acids, e.g., 1 or 2, e.g., conservative aminoacids, from the corresponding CDR of SEQ ID NO: 25 and each LC CDRdiffers by no more than 1, 2, 3, 4 or 5 amino acids, e.g., 1 or 2, e.g.,conservative amino acids, from the corresponding CDR of SEQ ID NO: 52.

In an embodiment, the antibody molecule comprises one or both of: aheavy chain variable region comprising at least 60, 70, 80, 85, 90, 95,98 or 99 percent homology with SEQ ID NO: 25; and a light chain variableregion comprising at least 60, 70, 80, 85, 90, 95, 98 or 99 percenthomology with SEQ ID NO: 52, wherein the antibody molecule comprises 1,2, 3, 4, 5, or all of: (i) a HC CDR1 comprising: S at the 1st positionand A at the 3^(rd) position in HC CDR1; (ii) a HC CDR2 comprising oneor both, e.g., one of: V at the 2^(nd) position; or N at the 7^(th)position and Q at the 16^(th) position in HC CDR2; (iii) a HC CDR3comprising: R at the 3^(rd) position (and optionally, L at the 3^(rd)position); (iv) a LC CDR1 comprising one or both of, e.g., one of: I atthe 3^(rd) position; or D at the 6th position in LC CDR1; (v) a LC CDR2comprising one, two, or three of, e.g., one of: G at the 2^(nd)position; Y at the 4^(th)position; or L at the 5^(th) position in LCCDR2; (vi) a LC CDR3 comprising: S at the 9^(th) position in LC CDR3.

In an embodiment, the antibody molecule, comprises: (a) a heavy chainimmunoglobulin variable region segment comprising SEQ ID NO: 25 (or asequence that differs by no more than 1, 2, 3, 4 or 5 amino acids, e.g.,conservative amino acids, therefrom); and (b) a light chain variableregion segment comprising SEQ ID NO: 52 (or a sequence that differs byno more than 1, 2, 3, 4 or 5 amino acids, e.g., conservative aminoacids, therefrom).

In an embodiment, the antibody molecule comprises one or both of: (a) aheavy chain immunoglobulin variable region segment comprising a CDR1comprising the sequence S-Y-A-M-H (SEQ ID NO: 68) (or a sequence thatdiffers by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2 amino acids,e.g., conservative amino acids, therefrom); a CDR2 comprising thesequence V-V-S-Y-D-G-N-Y-K-Y-Y-A-D-S-V-Q-G (SEQ ID NO: 69) (or asequence that differs by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2amino acids, e.g., conservative amino acids, therefrom); a CDR3comprising the sequence D-S-R-L-R-S-L-L-Y-F-E-W-L-S-Q-G-Y-F-N-P (SEQ IDNO: 70) (or a sequence that differs by no more than, 1, 2, 3, 4, or 5,e.g., 1 or 2 amino acids, e.g., conservative amino acids, therefrom);and (b) a light chain variable region segment comprising a CDR1comprising the sequence: Q-S-I-T-F-D-Y-K-N-Y-L-A (SEQ ID NO: 145) (or asequence that differs by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2amino acids, e.g., conservative amino acids, therefrom); a CDR2comprising the sequence W-G-S-Y-L-E-S (SEQ ID NO: 72) (or a sequencethat differs by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2 aminoacids, e.g., conservative amino acids, therefrom); a CDR3 comprising thesequence Q-Q-H-Y-R-T-P-P-S (SEQ ID NO: 73) (or a sequence that differsby no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2 amino acids, e.g.,conservative amino acids, therefrom).

In an embodiment, the antibody molecule comprises one or both of: a) LCCDR1-3, that collectively, differ from the Ab 044 LC CDR1-3 by no morethan, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, e.g., 1, 2, 3, or 4, aminoacids, e.g., conservative amino acids; and b) HC CDR1-3, thatcollectively, differ from the Ab 044 HC CDR1-3 by no more than, 1, 2, 3,4, 5, 6, 7, 8, 9, or 10, e.g., 1, 2, 3, or 4, amino acids, e.g.,conservative amino acids.

In an embodiment, the antibody molecule comprises one or both of: (a) aheavy chain immunoglobulin variable region segment comprising SEQ ID NO:25; and (b) a light chain variable region segment comprising SEQ ID NO:52.

In an embodiment, the antibody molecule comprises one or both of: (a) aheavy chain immunoglobulin variable region segment comprising a CDR1comprising the sequence S-Y-A-M-H (SEQ ID NO: 68) (or a sequence thatdiffers by no more than, 1, 2, or 3, e.g., 1 or 2, amino acids, e.g.,conservative amino acids, there from, optionally provided that at least1 or 2 of the highlighted residue are not changed, e.g., both S and Aare not changed); a CDR2 comprising the sequenceV-V-S-Y-D-G-N-Y-K-Y-Y-A-D-S-V-O-G (SEQ ID NO: 69) (or a sequence thatdiffers by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2 amino acids,e.g., conservative amino acids, therefrom, optionally provided that atleast 1, 2, or 3 of the highlighted residues are not changed, e.g., V orboth N and Q or all three of V, N, and Q are not changed); a CDR3comprising the sequence D-S-R-L-R-S-L-L-Y-F-E-W-L-S-Q-G-Y-F-N-P (SEQ IDNO: 70) (or a sequence that differs by no more than, 1, 2, 3, 4, or 5,e.g., 1 or 2 amino acids, e.g., conservative amino acids, therefrom,optionally provided that R is not changed); and (b) a light chainvariable region segment comprising a CDR1 comprising the sequence:Q-S-I-T-F-D-Y-K-N-Y-L-A (SEQ ID NO: 145) (or a sequence that differs byno more than, 1, 2, 3, 4, or 5, e.g., 1 or 2, amino acids, e.g.,conservative amino acids, therefrom, optionally provided that at leastfor 2 of the highlighted residues are not changed, e.g., I or D is notchanged); a CDR2 comprising the sequence W-G-S-Y-L-E-S (SEQ ID NO: 72)(or a sequence that differs by no more than, 1, 2, 3, 4, or 5, e.g., 1or 2, amino acids, e.g., conservative amino acids, therefrom, optionallyprovided that at least 1, 2 or 3 of the highlighted residues are notchanged, e.g., 1, 2 or all of G, Y, and L are not changed); a CDR3comprising the sequence Q-Q-H-Y-R-T-P-P-S (SEQ ID NO: 73) (or a sequencethat differs by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2, aminoacids, e.g., conservative amino acids, therefrom, optionally providedthat at least 1 or both of the highlighted residues are not changed,e.g., S is not changed). In an embodiment, a CDR of the light or heavychain includes one of the highlighted residues, or one of thehighlighted combinations of residues, for that CDR, (e.g., while otherresidues in that CDR might be changed, the highlighted residue orcombination of residues, are not changed). For example, in anembodiment, V or both N and Q, for heavy chain CDR2 are not changed.

In an embodiment, a CDR of the light chain and a CDR of the heavy chaineach includes one of the highlighted residues, or one of the highlightedcombinations of residues, for that CDR. In an embodiment, each of twoCDRs in the antibody molecule includes one of the highlighted residues,or one of the highlighted combinations of residues, for that CDR. In anembodiment, both are in the light chain. In an embodiment, both are inthe heavy chain. In an embodiment, each of the three CDRs in the heavychain includes one of the highlighted residues, or one of thehighlighted combinations of residues, for that CDR. In an embodiment,each of the three CDRs in the light chain includes one of thehighlighted residues, or one of the highlighted combinations ofresidues, for that CDR. In an embodiment, each of the six CDRs in theheavy and light chain includes one of the highlighted residues, or oneof the highlighted combinations of residues, for that CDR.

In an embodiment, the antibody molecule comprises one or more (e.g., 2,3, 4, 5, or all) of the following properties: (a) both S and A in HCCDR1 are unchanged; (b) V or both N and Q or all three of V, N, and Q inHC CDR2 are unchanged; (c) R in HC CDR3 is unchanged; (d) One or both ofI and D in LC CDR1 are unchanged; (e) 1, 2 or 3 of G, Y and L in LC CDR2are unchanged; or (f) S in LC CDR3 is unchanged. In an embodiment, theantibody molecule comprises 1, 2, 3, 4, 5, or all 6 properties selectedfrom (a) to (f). In an embodiment, the antibody molecule comprises aheavy chain having a one or more properties selected from (a), (b), and(c) and a light chain having one or more properties selected from (d),(e), and (f).

In an embodiment, the antibody molecule comprises one or both of: (a) aheavy chain immunoglobulin variable region segment comprising: a CDR1comprising the sequence S-Y-A-M-H (SEQ ID NO: 68); a CDR2 comprising thesequence V-V-S-Y-D-G-N-Y-K-Y-Y-A-D-S-V-Q-G (SEQ ID NO: 69); a CDR3comprising the sequence D-S-R-L-R-S-L-L-Y-F-E-W-L-S-Q-G-Y-F-N-P (SEQ IDNO: 70); and (b) a light chain variable region segment comprising a CDR1comprising the sequence Q-S-I-T-F-D-Y-K-N-Y-L-A (SEQ ID NO: 145); a CDR2comprising the sequence W-G-S-Y-L-E-S (SEQ ID NO:72); and a CDR3comprising the sequence Q-Q-H-Y-R-T-P-P-S (SEQ ID NO: 73).

In an embodiment, the antibody molecule comprises one or both of: a) oneor more framework regions (FRs) from SEQ ID NO: 25 e.g., the antibodymolecule comprises one or more or all of FR1, FR2, FR3, or FR4, orsequences that differ individually, or collectively, by no more than 1,2, 3, 4, of 5 amino acid residues, e.g., conservative residues, from SEQID NO: 25; and b) one or more framework regions (FRs) from SEQ ID NO:52. For example, the antibody molecule comprises one or more or all ofFR1, FR2, FR3, or FR4, or sequences that differ individually, orcollectively, by no more than 1, 2, 3, 4, of 5 amino acid residues,e.g., conservative residues, from SEQ ID NO: 52.

In an embodiment, the antibody molecule comprises: (a) a heavy chainimmunoglobulin variable region segment that further comprises one ormore or all of: an FR1 comprising the sequenceQ-V-Q-L-L-E-T-G-G-G-L-V-K-P-G-Q-S-L-K-L-S-C-A-A-S-G-F-T-F-T (SEQ ID NO:74) (or a sequence that differs by no more than, 1, 2, 3, 4, or 5, e.g.,1 or 2, amino acids, e.g., conservative amino acids, therefrom,optionally provided that T is not changed); an FR2 comprising thesequence W-V-R-Q-P-P-G-K-G-L-E-W-V-A (SEQ ID NO: 75) (or a sequence thatdiffers by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2, amino acids,e.g., conservative amino acids, therefrom, optionally provided that W isnot changed, or that if changed, is other than R); an FR3 comprising thesequence R-F-T-I-S-R-D-N-S-K-N-T-L-Y-L-Q-M-N-S-L-R-A-E-D-T-A-V-Y-Y-C-A-K(SEQ ID NO: 76) (or a sequence that differs by no more than, 1, 2, 3, 4,or 5, e.g., 1 or 2, amino acids, e.g., conservative amino acids,therefrom, optionally provided that one, two or three of I, R, or L isnot changed, or that if I is changed it is other than G, if R is changedit is other than P. or if L is changed it is other than A); and an FR4comprising the sequence W-G-Q-G-T-T-L-T-V-S-S (SEQ ID NO: 77) (or asequence that differs by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2,amino acids, e.g., conservative amino acids, therefrom) orW-G-Q-G-T-T-V-T-V-S-S (SEQ ID NO: 171) (or a sequence that differs by nomore than, 1, 2, 3, 4, or 5, e.g., 1 or 2, amino acids, e.g.,conservative amino acids, therefrom); and (b) a light chainimmunoglobulin variable region segment comprising one or more or all of:an FR1 comprising the sequenceD-I-Q-M-T-Q-S-P-S-S-L-S-A-S-V-G-D-R-V-T-I-T-C-R-S-S (SEQ ID NO: 78) (ora sequence that differs by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2,amino acids, e.g., conservative amino acids, therefrom, optionallyprovided that R is not changed); an FR2 comprising the sequenceW-Y-Q-Q-K-P-G-K-A-P-K-L-L-I-Y (SEQ ID NO: 79) (or a sequence thatdiffers by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2 amino acids,e.g., conservative amino acids, therefrom); an FR3 comprising thesequence G-V-P-S-R-F-S-G-S-G-S-G-T-D-F-T-L-T-I-S-S-L-Q-P-E-D-F-A-T-Y-Y-C(SEQ ID NO: 80) (or a sequence that differs by no more than, 1, 2, 3, 4,or 5, e.g., 1 or 2 amino acids, e.g., conservative amino acids,therefrom, optionally provided that C is not changed, or if changed, isother than P); and an FR4 comprising the sequence F-G-Q-G-T-K-V-E-I-K(SEQ ID NO: 81) (or a sequence that differs by no more than, 1, 2, 3, 4,or 5, e.g., 1 or 2 amino acids, e.g., conservative amino acids,therefrom). In an embodiment, a FR of the light or heavy chain includesone of the highlighted residues, or one of the highlighted combinationsof residues, for that FR, (e.g., while other residues in that FR mightbe changed, the highlighted residue or combination of residues, are notchanged). For example, in an embodiment, one, two or three of I, R, or Lfor heavy chain FR3 is not changed.

In an embodiment, a FR of the light chain and a FR of the heavy chaineach includes one of the highlighted residues, or one of the highlightedcombinations of residues, for that FR. In an embodiment, each of two FRsin the antibody molecule includes one of the highlighted residues, orone of the highlighted combinations of residues, for that FR. In anembodiment, both are in the light chain. In an embodiment, both are inthe heavy chain. In an embodiment, each of FR2 and FR3 in the heavychain includes one of the highlighted residues, or one of thehighlighted combinations of residues, for that FR. In an embodiment,each of FR1 and FR2 in the heavy and light chain includes one of the highlighted residues for that FR. In an embodiment, all of thehighlighted residues in heavy chain FR1-4 are unchanged. In anembodiment, all of the highlighted residues in light chain FR1-4 areunchanged. In an embodiment, all of the highlighted residues in bothheavy and light chain FR1-4 are unchanged.

In an embodiment, sequence of FR1 of the heavy chain variable regionsegment is Q-V-Q-L-L-E-T-G-G-G-L-V-K-P-G-Q-S-L-K-L-S-C-A-A-S-G-F-T-F-T(SEQ ID NO: 74). In an embodiment, sequence of FR1 of the heavy chainvariable region segment isE-V-Q-L-L-E-S-G-G-G-L-V-K-P-G-Q-S-L-K-L-S-C-A-A-S-G-F-T-F-T (SEQ ID NO:183).

In another embodiment, the antibody molecule comprises a structural orfunctional property of Ab 069.

In an embodiment, the antibody molecule competes with a referenceantibody molecule, e.g., an antibody molecule described herein, forbinding to a substrate, e.g., an HA. The reference antibody molecule canbe: a) an antibody molecule comprising: i) a heavy chain immunoglobulinvariable region segment comprising a CDR1 comprising the sequenceS-Y-A-M-H (SEQ ID NO: 68); a CDR2 comprising the sequenceV-V-S-Y-D-G-N-Y-K-Y-Y-A-D-S-V-Q-G (SEQ ID NO: 69); and a CDR3 comprisingthe sequence D-S-R-L-R-S-L-L-Y-F-E-W-L-S-Q-G-Y-F-N-P (SEQ ID NO: 70);and ii) a light chain variable region segment comprising: a CDR1comprising the sequence Q-S-I-T-F-E-Y-K-N-Y-L-A (SEQ ID NO:172); a CDR2comprising the sequence W-G-S-Y-L-E-S (SEQ ID NO:72); and a CDR3comprising the sequence Q-Q-H-Y-R-T-P-P-S (SEQ ID NO: 73); b) anantibody molecule comprises one or both of: (i) a heavy chainimmunoglobulin variable region segment comprising SEQ ID NO: 25; and(ii) a light chain variable region segment comprising SEQ ID NO:155; orc) Ab 069.

In an embodiment, the antibody molecule binds to the same epitope, or aportion thereof, on HA, as does a reference antibody molecule, e.g. anantibody molecule disclosed herein. The reference antibody molecule canbe: a) an antibody molecule comprising: i) a heavy chain immunoglobulinvariable region segment comprising a CDR1 comprising the sequenceS-Y-A-M-H (SEQ ID NO: 68); a CDR2 comprising the sequenceV-V-S-Y-D-G-N-Y-K-Y-Y-A-D-S-V-Q-G (SEQ ID NO: 69); and a CDR3 comprisingthe sequence D-S-R-L-R-S-L-L-Y-F-E-W-L-S-Q-G-Y-F-N-P (SEQ ID NO: 70);and ii) a light chain variable region segment comprising: a CDR1comprising the sequence Q-S-I-T-F-E-Y-K-N-Y-L-A (SEQ ID NO: 172); a CDR2comprising the sequence W-G-S-Y-L-E-S (SEQ ID NO:72); and a CDR3comprising the sequence Q-Q-H-Y-R-T-P-P-S (SEQ ID NO: 73); b) anantibody molecule comprises one or both of: (i) a heavy chainimmunoglobulin variable region segment comprising SEQ ID NO: 25; and(ii) a light chain variable region segment comprising SEQ ID NO: 155; orc) Ab 069.

In an embodiment the antibody molecule, comprises one or both of: aheavy chain variable region comprising at least 60, 70, 80, 85, 90, 95,98 or 99 percent homology with SEQ ID NO: 25; and a light chain variableregion comprising at least 60, 70, 80, 85, 90, 95, 98 or 99 percenthomology with SEQ ID NO: 155.

In an embodiment the antibody molecule, comprises one or both of: aheavy chain variable region comprising at least 60, 70, 80, 85, 90, 95,98 or 99 percent homology with SEQ ID NO: 25; and a light chain variableregion comprising at least 60, 70, 80, 85, 90, 95, 98 or 99 percenthomology with SEQ ID NO: 155, wherein each HC CDR differs by no morethan 1, 2, 3, 4 or 5 amino acids, e.g., 1 or 2, e.g., conservative aminoacids, from the corresponding CDR of SEQ ID NO: 25 and each LC CDRdiffers by no more than 1, 2, 3, 4 or 5 amino acids, e.g., 1 or 2, e.g.,conservative amino acids, from the corresponding CDR of SEQ ID NO: 155.

In an embodiment the antibody molecule, comprises one or both of: aheavy chain variable region comprising at least 60, 70, 80, 85, 90, 95,98 or 99 percent homology with SEQ ID NO: 25; and a light chain variableregion comprising at least 60, 70, 80, 85, 90, 95, 98 or 99 percenthomology with SEQ ID NO: 155, wherein the antibody molecule comprises 1,2, 3, 4, 5, or all of: (i) a HC CDR1 comprising: S at the 1st positionand A at the 3^(rd) position in HC CDR1; (ii) a HC CDR2 comprising oneor both, e.g., one of: V at the 2nd position; or N at the 7^(th)position and Q at the 16^(th) position in HC CDR2; (iii) a HC CDR3comprising: R at the 3^(rd) position (and optionally, L at the 3^(rd)position); (iv) a LC CDR1 comprising one or both of, e.g., one of: I atthe 3^(rd) position; or E at the 6^(th) position in LC CDR1; (v) a LCCDR2 comprising one, two or three of, e.g., one of: G at the 2^(nd)position; Y at the 4^(th) position; or L at the 5^(th) position in LCCDR2; (vi) a LC CDR3 comprising: S at the 9^(th) position in LC CDR3.

In an embodiment, the antibody molecule, comprises one or both of: (a) aheavy chain immunoglobulin variable region segment comprising SEQ ID NO:25 (or a sequence that differs by no more than 1, 2, 3, 4 or 5 aminoacids, e.g., conservative amino acids, therefrom); and (b) a light chainvariable region segment comprising SEQ ID NO: 155 (or a sequence thatdiffers by no more than 1, 2, 3, 4 or 5 amino acids, e.g., conservativeamino acids, therefrom).

In an embodiment, the antibody molecule comprises one or both of: (a) aheavy chain immunoglobulin variable region segment comprising SEQ ID NO:25; and (b) a light chain variable region segment comprising SEQ ID NO:155.

In an embodiment, the antibody molecule comprises one or both of: (a) aheavy chain immunoglobulin variable region segment comprising a CDR1comprising the sequence S-Y-A-M-H (SEQ ID NO: 68) (or a sequence thatdiffers by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2 amino acids,e.g., conservative amino acids, therefrom); a CDR2 comprising thesequence V-V-S-Y-D-G-N-Y-K-Y-Y-A-D-S-V-Q-G (SEQ ID NO: 69) (or asequence that differs by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2amino acids, e.g., conservative amino acids, therefrom); a CDR3comprising the sequence D-S-R-L-R-S-L-L-Y-F-E-W-L-S-Q-G-Y-F-N-P (SEQ IDNO: 70) (or a sequence that differs by no more than, 1, 2, 3, 4, or 5,e.g., 1 or 2 amino acids, e.g., conservative amino acids, therefrom);and (b) a light chain variable region segment comprising a CDR1comprising the sequence: Q-S-I-T-F-E-Y-K-N-Y-L-A (SEQ ID NO: 172) or asequence that differs by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2amino acids, e.g., conservative amino acids, therefrom); a CDR2comprising the sequence W-G-S-Y-L-E-S (SEQ ID NO: 72) (or a sequencethat differs by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2 aminoacids, e.g., conservative amino acids, therefrom); a CDR3 comprising thesequence Q-Q-H-Y-R-T-P-P-S (SEQ ID NO: 73) (or a sequence that differsby no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2 amino acids, e.g.,conservative amino acids, therefrom).

In an embodiment, the antibody molecule comprises one or both of: a) LCCDR1-3, that collectively, differ from the Ab 069 LC CDR1-3 by no morethan, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, e.g., 1, 2, 3, or 4, aminoacids, e.g., conservative amino acids; and b) HC CDR1-3, thatcollectively, differ from the Ab 069 HC CDR1-3 by no more than, 1, 2, 3,4, 5, 6, 7, 8, 9, or 10, e.g., 1, 2, 3, or 4, amino acids, e.g.,conservative amino acids.

In an embodiment, the antibody molecule comprises one or both of: (a) aheavy chain immunoglobulin variable region segment comprising a CDR1comprising the sequence S-Y-A-M-H (SEQ ID NO: 68) (or a sequence thatdiffers by no more than, 1, 2, or 3, e.g., 1 or 2 amino acids, e.g.,conservative amino acids, therefrom, optionally provided that at least 1or 2 of the highlighted residues are not changed, e.g., both S and A arenot changed); a CDR2 comprising the sequenceV-V-S-Y-D-G-N-Y-K-Y-Y-A-D-S-V-Q-G (SEQ ID NO: 69) (or a sequence thatdiffers by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2 amino acids,e.g., conservative amino acids, therefrom, optionally provided that atleast 1, 2, or 3 of the highlighted residues are not changed, e.g., V orboth N and Q or all three of V, N, and Q are not changed); a CDR3comprising the sequence D-S-R-L-R-S-L-L-Y-F-E-W-L-S-Q-G-Y-F-N-P (SEQ IDNO: 70) (or a sequence that differs by no more than, 1, 2, 3, 4, or 5,e.g., 1 or 2 amino acids, e.g., conservative amino acids, therefromoptionally provided that, R is not changed); and (b) a light chainvariable region segment comprising a CDR1 comprising the sequence:Q-S-I-T-F-E-Y-K-N-Y-L-A (SEQ ID NO: 172) or a sequence that differs byno more than, 1, 2, 3, 4, or 5, e.g., 1 or 2 amino acids, e.g.,conservative amino acids, therefrom, optionally provided that at least 1or 2 of the highlighted residues are not changed, e.g., I or E is notchanged); a CDR2 comprising the sequence W-G-S-Y-L-E-S (SEQ ID NO: 72)(or a sequence that differs by no more than, 1, 2, 3, 4, or 5, e.g., 1or 2 amino acids, e.g., conservative amino acids, therefrom, optionallyprovided that at least 1, 2, or 3 of the highlighted residues are notchanged, e.g., 1, 2 or all of G, Y, and L are not changed); a CDR3comprising the sequence Q-Q-H-Y-R-T-P-P-S (SEQ ID NO: 73) (or a sequencethat differs by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2 aminoacids, e.g., conservative amino acids, therefrom, optionally providedthat, at least one or both of the highlighted residues are not changed,e.g., S is not changed).

In an embodiment, a CDR of the light or heavy chain includes one of thehighlighted residues, or one of the highlighted combinations ofresidues, for that CDR (e.g., while other residues in that CDR might bechanged, the highlighted residue or combination of residues, are notchanged). In an embodiment, a CDR of the light and a CDR of the heavychain each includes one of the highlighted residues, or one of thehighlighted combinations of residues, for that CDR. In an embodiment,each of two CDRs in the antibody molecule includes one of thehighlighted residues, or one of the highlighted combinations ofresidues, for that CDR. In an embodiment, both are in the light chain.In an embodiment, both are in the heavy chain. In an embodiment, each ofthe three CDRs in the heavy chain includes one of the highlightedresidues, or one of the highlighted combinations of residues, for thatCDR. In an embodiment, each of the three CDRs in the light chainincludes one of the highlighted residues, or one of the highlightedcombinations of residues, for that CDR. In an embodiment, each of thesix CDRs in the heavy and light chain includes one of the highlightedresidues, or one of the highlighted combinations of residues, for thatCDR.

In an embodiment, the antibody molecule comprises one or more (e.g., 2,3, 4, 5, or all) of the following properties: (a) both S and A in HCCDR1 are unchanged; (b) V or both N and Q or all three of V, N, and Q inHC CDR2 are unchanged; (c) R in HC CDR3 is unchanged; (d) one or both ofI and E in LC CDR1 are unchanged; (e) 1, 2 or 3 of G, Y, and L in LCCDR2 are unchanged; or (f) S in LC CDR3 is unchanged. In an embodimentthe antibody molecule comprises 1, 2, 3, 4, 5, or all 6 propertiesselected from (a) to (f). In an embodiment, the antibody moleculecomprises a heavy chain having a one or more properties selected from(a), (b), and (c) and a light chain having one or more propertiesselected from (d), (e), and (f).

In an embodiment, the antibody molecule comprises one or both of: (a) aheavy chain immunoglobulin variable region segment comprising: a CDR1comprising the sequence S-Y-A-M-H (SEQ ID NO: 68); a CDR2 comprising thesequence V-V-S-Y-D-G-N-Y-K-Y-Y-A-D-S-V-Q-G (SEQ ID NO: 69); a CDR3comprising the sequence D-S-R-L-R-S-L-L-Y-F-E-W-L-S-Q-G-Y-F-N-P (SEQ IDNO: 70); and (b) a light chain variable region segment comprising a CDR1comprising the sequence Q-S-I-T-F-E-Y-K-N-Y-L-A (SEQ ID NO: 172); a CDR2comprising the sequence W-G-S-Y-L-E-S (SEQ ID NO: 72); and a CDR3comprising the sequence Q-Q-H-Y-R-T-P-P-S (SEQ ID NO: 73).

In an embodiment, the antibody molecule comprises one or both of: a) oneor more framework regions (FRs) from SEQ ID NO: 25, e.g., one or more orall of FR1, FR2, FR3, or FR4, or sequences that differ individually, orcollectively, by no more than 1, 2, 3, 4, of 5 amino acid residues,e.g., conservative residues, from SEQ ID NO: 25; and b) one or moreframework regions (FRs) from SEQ ID NO: 155, e.g., one or more or all ofFR1, FR2, FR3, or FR4, or sequences that differ individually, orcollectively, by no more than 1, 2, 3, 4, of 5 amino acid residues,e.g., conservative residues, from SEQ ID NO: 155.

In an embodiment, the antibody molecule comprises: (a) a heavy chainimmunoglobulin variable region segment that further comprises one ormore or all of: an FR1 comprising the sequenceQ-V-Q-L-L-E-T-G-G-G-L-V-K-P-G-Q-S-L-K-L-S-C-A-A-S-G-F-T-F-T (SEQ IDNO:74) (or a sequence that differs by no more than, 1, 2, 3, 4, or 5,e.g., 1 or 2, amino acids, e.g., conservative amino acids, therefrom,optionally provided that T is not changed); an FR2 comprising thesequence W-V-R-Q-P-P-G-K-G-L-E-W-V-A (SEQ ID NO: 75) (or a sequence thatdiffers by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2, amino acids,e.g., conservative amino acids, therefrom, optionally provided that W isnot changed, or that if changed, is other than R); an FR3 comprising thesequence R-F-T-I-S-R-D-N-S-K-N-T-L-Y-L-Q-M-N-S-L-R-A-E-D-T-A-V-Y-Y-C-A-K(SEQ ID NO: 76) (or a sequence that differs by no more than, 1, 2, 3, 4,or 5, e.g., 1 or 2, amino acids, e.g., conservative amino acids,therefrom, optionally provided that one, two or three of I, R, or L isnot changed, or that if I is changed it is other than G, if R is changedit is other than P. or if L is changed it is other than A); and (b) thelight chain immunoglobulin variable region segment comprises one or moreor all of an FR1 comprising the sequenceD-I-Q-M-T-Q-S-P-S-S-L-S-A-S-V-G-D-R-V-T-I-T-C-R-S-S (SEQ ID NO: 78) (ora sequence that differs by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2amino acids, e.g., conservative amino acids, therefrom, optionallyprovided that R is not changed); an FR2 comprising the sequenceW-Y-Q-Q-K-P-G-K-A-P-K-L-L-I-Y (SEQ ID NO: 79) (or a sequence thatdiffers by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2 amino acids,e.g., conservative amino acids, therefrom); an FR3 comprising thesequence G-V-P-S-R-F-S-G-S-G-S-G-T-D-F-T-L-T-I-S-S-L-Q-P-E-D-F-A-T-Y-Y-C(SEQ ID NO: 80) (or a sequence that differs by no more than, 1, 2, 3, 4,or 5, e.g., 1 or 2 amino acids, e.g., conservative amino acids,therefrom, optionally provided that C is not changed, or if changed, isother than P); and an FR4 comprising the sequence F-G-Q-G-T-K-V-E-I-K(SEQ ID NO: 81) (or a sequence that differs by no more than, 1, 2, 3, 4,or 5, e.g., 1 or 2 amino acids, e.g., conservative amino acids,therefrom).

In an embodiment a FR of the light or heavy chain includes one of thehighlighted residues, or one of the highlighted combinations ofresidues, for that FR (e.g., while other residues in that FR might bechanged, the highlighted residue or combination of residues, are notchanged). For example, in an embodiment, one, two or three of I, R, or Lfor heavy chain FR3 is not changed.

In an embodiment, a FR of the light chain and a FR of the heavy chaineach includes one of the highlighted residues, or one of the highlightedcombinations of residues, for that FR. In an embodiment, each of two FRsin the antibody molecule includes one of the highlighted residues, orone of the highlighted combinations of residues, for that FR. In anembodiment, both are in the light chain. In an embodiment, both are inthe heavy chain. In an embodiment, each of FR2 and FR3 in the heavychain includes one of the highlighted residues, or one of thehighlighted combinations of residues, for that FR. In an embodiment,each of FR1 and FR2 in the heavy and light chain includes one of the highlighted residues for that FR. In an embodiment, all of thehighlighted residues in heavy chain FR1-4 are unchanged. In anembodiment, all of the highlighted residues in light chain FR1-4 areunchanged. In an embodiment, all of the highlighted residues in bothheavy and light chain FR1-4 are unchanged.

In another embodiment, the antibody molecule comprises a structural orfunctional property of Ab 032.

In an embodiment, the antibody molecule competes with a referenceantibody molecule, e.g., an antibody molecule described herein, forbinding to a substrate, e.g., an HA. The reference antibody molecule canbe: a) an antibody molecule comprising: i) a heavy chain immunoglobulinvariable region segment comprising a CDR1 comprising the sequenceS-Y-A-M-H (SEQ ID NO: 68); a CDR2 comprising the sequenceV-V-S-Y-D-G-N-Y-K-Y-Y-A-D-S-V-Q-G (SEQ ID NO: 69); and a CDR3 comprisingthe sequence D-S-R-L-R-S-L-L-Y-F-E-W-L-S-Q-G-Y-F-N-P (SEQ ID NO: 70);and ii) a light chain variable region segment comprising: a CDR1comprising the sequence Q-S-I-T-F-N-Y-K-N-Y-L-A (SEQ ID NO: 71); a CDR2comprising the sequence W-G-S-Y-L-E-S (SEQ ID NO: 72); and a CDR3comprising the sequence Q-Q-H-Y-R-T-P-P-S (SEQ ID NO: 73); b) anantibody molecule comprises one or both of: (i) a heavy chainimmunoglobulin variable region segment comprising SEQ ID NO: 25; and(ii) a light chain variable region segment comprising SEQ ID NO:45; orc) Ab 032.

In an embodiment, the antibody molecule binds to the same epitope, or aportion thereof, on HA, as does a reference antibody molecule, e.g. anantibody molecule disclosed herein. The reference antibody molecule canbe: a) an antibody molecule comprising: i) a heavy chain immunoglobulinvariable region segment comprising a CDR1 comprising the sequenceS-Y-A-M-H (SEQ ID NO: 68); a CDR2 comprising the sequenceV-V-S-Y-D-G-N-Y-K-Y-Y-A-D-S-V-Q-G (SEQ ID NO: 69); and a CDR3 comprisingthe sequence D-S-R-L-R-S-L-L-Y-F-E-W-L-S-Q-G-Y-F-N-P (SEQ ID NO: 70);and ii) a light chain variable region segment comprising: a CDR1comprising the sequence Q-S-I-T-F-N-Y-K-N-Y-L-A (SEQ ID NO: 71); a CDR2comprising the sequence W-G-S-Y-L-E-S (SEQ ID NO: 72); and a CDR3comprising the sequence Q-Q-H-Y-R-T-P-P-S (SEQ ID NO: 73); b) anantibody molecule comprises one or both of: (i) a heavy chainimmunoglobulin variable region segment comprising SEQ ID NO: 25; and(ii) a light chain variable region segment comprising SEQ ID NO: 45; orc) Ab 032.

In an embodiment, the antibody molecule comprises one or both of: aheavy chain variable region comprising least 60, 70, 80, 85, 90, 95, 98or 99 percent homology with SEQ ID NO: 25; or a light chain variableregion comprising least 60, 70, 80, 85, 90, 95, 98 or 99 percenthomology with SEQ ID NO: 45.

In an embodiment, the antibody molecule comprises one or both of: aheavy chain variable region comprising least 60, 70, 80, 85, 90, 95, 98or 99 percent homology with SEQ ID NO: 25; and a light chain variableregion comprising least 60, 70, 80, 85, 90, 95, 98 or 99 percenthomology with SEQ ID NO: 45, wherein each HC CDR differs by no more than1, 2, 3, 4 or 5 amino acids, e.g., 1 or 2, e.g., conservative aminoacids, from the corresponding CDR of SEQ ID NO: 25 and each LC CDRdiffers by no more than 1, 2, 3, 4 or 5 amino acids, e.g., 1 or 2, e.g.,conservative amino acids, from the corresponding CDR of SEQ ID NO: 45.

In an embodiment, the antibody molecule comprises one or both of: aheavy chain variable region comprising least 60, 70, 80, 85, 90, 95, 98or 99 percent homology with SEQ ID NO: 25; and a light chain variableregion comprising least 60, 70, 80, 85, 90, 95, 98 or 99 percenthomology with SEQ ID NO: 45, wherein the antibody molecule comprises 1,2, 3, 4, 5, or all of: (i) a HC CDR1 comprising: S at the 1st positionand A at the 3^(rd) position in HC CDR1; (ii) a HC CDR2 comprising oneor both, e.g., one of: V at the 2^(nd) position; or N at the 7^(th)position and Q at the 16^(th) position in HC CDR2; (iii) a HC CDR3comprising: R at the 3^(rd) position (and optionally, L at the 3^(rd)position); (iv) a LC CDR1 comprising: I at the 3^(rd) position; (v) a LCCDR2 comprising one, two, or three of, e.g., one of: G at the 2^(nd)position; Y at the 4^(th) position; or L at the 5^(th) position in LCCDR2; (vi) a LC CDR3 comprising: S at the 9^(th) position in LC CDR3.

In an embodiment, the antibody molecule comprises one or both of: (a) aheavy chain immunoglobulin variable region segment comprising SEQ IDNO:25 (or a sequence that differs by no more than 1, 2, 3, 4 or 5 aminoacids, e.g., conservative amino acids, therefrom); and (b) a light chainvariable region segment comprising SEQ ID NO:155 (or a sequence thatdiffers by no more than 1, 2, 3, 4 or 5 amino acids, e.g., conservativeamino acids, therefrom).

In an embodiment, the antibody molecule comprises one or both of: (a) aheavy chain immunoglobulin variable region segment comprising SEQ ID NO:25; and (b) a light chain variable region segment comprising SEQ ID NO:155.

In an embodiment, the antibody molecule comprises one or both of: (a) aheavy chain immunoglobulin variable region segment comprising a CDR1comprising the sequence S-Y-A-M-H (SEQ ID NO: 68) (or a sequence thatdiffers by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2 amino acids,e.g., conservative amino acids, therefrom); a CDR2 comprising thesequence V-V-S-Y-D-G-N-Y-K-Y-Y-A-D-S-V-Q-G (SEQ ID NO: 69) (or asequence that differs by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2amino acids, e.g., conservative amino acids, therefrom); a CDR3comprising the sequence D-S-R-L-R-S-L-L-Y-F-E-W-L-S-Q-G-Y-F-N-P (SEQ IDNO: 70) (or a sequence that differs by no more than, 1, 2, 3, 4, or 5,e.g., 1 or 2 amino acids, e.g., conservative amino acids, therefrom);and (b) a light chain variable region segment comprising a CDR1comprising the sequence: Q-S-I-T-F N-Y-K-N-Y-L-A (SEQ ID NO: 71) (or asequence that differs by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2amino acids, e.g., conservative amino acids, therefrom); a CDR2comprising the sequence W-G-S-Y-L-E-S (SEQ ID NO: 72) (or a sequencethat differs by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2 aminoacids, e.g., conservative amino acids, therefrom); a CDR3 comprising thesequence Q-Q-H-Y-R-T-P-P-S (SEQ ID NO: 73) (or a sequence that differsby no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2 amino acids, e.g.,conservative amino acids, therefrom).

In an embodiment, the antibody molecule comprises one or both of: a) LCCDR1-3, that collectively, differ from the Ab 032 LC CDR1-3 by no morethan, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, e.g., 1, 2, 3, or 4, aminoacids, e.g., conservative amino acids; and b) HC CDR1-3, thatcollectively, differ from the Ab 032 HC CDR1-3 by no more than, 1, 2, 3,4, 5, 6, 7, 8, 9, or 10, e.g., 1, 2, 3, or 4, amino acids, e.g.,conservative amino acids.

In an embodiment, the antibody molecule comprises one or both of: (a) aheavy chain immunoglobulin variable region segment comprising a CDR1comprising the sequence S-Y-A-M-H (SEQ ID NO: 68) (or a sequence thatdiffers by no more than, 1, 2, or 3, e.g., 1 or 2 amino acids, e.g.,conservative amino acids, therefrom, optionally provided that at least 1or 2 of the highlighted residues are not changed, e.g., both S and A arenot changed); a CDR2 comprising the sequenceV-V-S-Y-D-G-N-Y-K-Y-Y-A-D-S-V-Q-G (SEQ ID NO: 69) (or a sequence thatdiffers by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2 amino acids,e.g., conservative amino acids, therefrom, provided that, e.g., at least1, 2, or 3 of the highlighted residues are not changed, e.g., V or bothN and Q or all three of V, N, and Q are not changed); a CDR3 comprisingthe sequence D-S-R-L-R-S-L-L-Y-F-E-W-L-S-Q-G-Y-F-N-P (SEQ ID NO: 70) (ora sequence that differs by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2amino acids, e.g., conservative amino acids, therefrom, optionallyprovided that R is not changed); and (b) a light chain variable regionsegment comprising a CDR1 comprising the sequence:Q-S-I-T-F-N-Y-K-N-Y-L-A (SEQ ID NO: 71) or a sequence that differs by nomore than, 1, 2, 3, 4, or 5, e.g., 1 or 2 amino acids, e.g.,conservative amino acids, therefrom, optionally provided that at least 1or 2 of the highlighted residues are not changed, e.g., I is notchanged); a CDR2 comprising the sequence W-G-S-Y-L-E-S (SEQ ID NO:72)(or a sequence that differs by no more than, 1, 2, 3, 4, or 5, e.g., 1or 2 amino acids, e.g., conservative amino acids, therefrom, optionallyprovided that at least 1, 2, or 3 of the highlighted residues are notchanged, e.g., 1, 2 or all of G, Y, and L are not changed); a CDR3comprising the sequence Q-Q-H-Y-R-T-P-P-S (SEQ ID NO: 73) (or a sequencethat differs by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2 aminoacids, e.g., conservative amino acids, therefrom, optionally providedthat at least one or both of the highlighted residues are not changed,e.g., S is not changed).

In an embodiment, a CDR of the light or heavy chain includes one of thehighlighted residues, or one of the highlighted combinations ofresidues, for that CDR (e.g., while other residues in that CDR might bechanged, the highlighted residue or combination of residues, are notchanged). In an embodiment, a CDR of the light and a CDR of the heavychain each includes one of the highlighted residues, or one of thehighlighted combinations of residues, for that CDR. In an embodiment,each of two CDRs in the antibody molecule includes one of thehighlighted residues, or one of the highlighted combinations ofresidues, for that CDR. In an embodiment, both are in the light chain.In an embodiment, both are in the heavy chain. In an embodiment each ofthe three CDRs in the heavy chain includes one of the highlightedresidues, or one of the highlighted combinations of residues, for thatCDR. In an embodiment, each of the three CDRs in the light chainincludes one of the highlighted residues, or one of the highlightedcombinations of residues, for that CDR. In an embodiment, each of thesix CDRs in the heavy and light chain includes one of the highlightedresidues, or one of the highlighted combinations of residues, for thatCDR.

In an embodiment, the antibody molecule comprises one or more (e.g., 2,3, 4, 5, or all) of the following properties: (a) both S and A in HCCDR1 are unchanged; (b) V or both N and Q or all three of V, N, and Q inHC CDR2 are unchanged; (c) R in HC CDR3 is unchanged; (d) I in LC CDR1is unchanged; (e) 1, 2 or 3 of G, Y, and L in LC CDR2 are unchanged; or(f) S in LC CDR3 is unchanged. In an embodiment, the antibody moleculecomprises 1, 2, 3, 4, 5, or all 6 properties selected from (a) to (f).In an embodiment, the antibody molecule comprises a heavy chain having aone or more properties selected from (a), (b), and (c) and a light chainhaving one or more properties selected from (d), (e), and (f).

In an embodiment, the antibody molecule comprises one or both of: (a) aheavy chain immunoglobulin variable region segment comprising: a CDR1comprising the sequence S-Y-A-M-H (SEQ ID NO: 68); a CDR2 comprising thesequence V-V-S-Y-D-G-N-Y-K-Y-Y-A-D-S-V-Q-G (SEQ ID NO: 69); a CDR3comprising the sequence D-S-R-L-R-S-L-L-Y-F-E-W-L-S-Q-G-Y-F-N-P (SEQ IDNO: 70); and (b) a light chain variable region segment comprising a CDR1comprising the sequence Q-S-I-T-F-N-Y-K-N-Y-L-A (SEQ ID NO: 71); a CDR2comprising the sequence W-G-S-Y-L-E-S (SEQ ID NO:72); and a CDR3comprising the sequence Q-Q-H-Y-R-T-P-P-S (SEQ ID NO:73).

In an embodiment, the antibody molecule comprises one or both of: a) oneor more framework regions (FRs) from SEQ ID NO: 25. For example, theantibody molecule comprises one or more or all of FR1, FR2, FR3, or FR4,or sequences that differ individually, or collectively, by no more than1, 2, 3, 4, of 5 amino acid residues, e.g., conservative residues, fromSEQ ID NO: 25; and b) one or more framework regions (FRs) from SEQ IDNO: 45. For example, the antibody molecule comprises one or more or allof FR1, FR2, FR3, or FR4, or sequences that differ individually, orcollectively, by no more than 1, 2, 3, 4, of 5 amino acid residues,e.g., conservative residues, from SEQ ID NO: 45.

In an embodiment, the antibody molecule comprises: (a) a heavy chainimmunoglobulin variable region segment that further comprises one ormore or all of: an FR1 comprising the sequenceQ-V-Q-L-L-E-T-G-G-G-L-V-K-P-G-Q-S-L-K-L-S-C-A-A-S-G-F-T-F-T (SEQ ID NO:74) (or a sequence that differs by no more than, 1, 2, 3, 4, or 5, e.g.,1 or 2, amino acids, e.g., conservative amino acids, therefrom,optionally provided that T is not changed); an FR2 comprising thesequence W-V-R-Q-P-P-G-K-G-L-E-W-V-A (SEQ ID NO: 75) (or a sequence thatdiffers by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2, amino acids,e.g., conservative amino acids, therefrom, optionally provided that W isnot changed, or that if changed, is other than R); an FR3 comprising thesequence R-F-T-I-S-R-D-N-S-K-N-T-L-Y-L-Q-M-N-S-L-R-A-E-D-T-A-V-Y-Y-C-A-K(SEQ ID NO: 76) (or a sequence that differs by no more than, 1, 2, 3, 4,or 5, e.g., 1 or 2, amino acids, e.g., conservative amino acids,therefrom, optionally provided that one, two or three of I, R, or L isnot changed, or that if I is changed it is other than G, if R is changedit is other than P. or if L is changed it is other than A); and an FR4comprising the sequence W-G-Q-G-T-T-L-T-V-S-S (SEQ ID NO: 77) (or asequence that differs by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2amino acids, e.g., conservative amino acids, therefrom) orW-G-Q-G-T-T-V-T-V-S-S (SEQ ID NO:171) (or a sequence that differs by nomore than, 1, 2, 3, 4, or 5, e.g., 1 or 2 amino acids, e.g.,conservative amino acids, therefrom); and (b) the light chainimmunoglobulin variable region segment comprises one or more or all ofan FR1 comprising the sequenceD-I-Q-M-T-Q-S-P-S-S-L-S-A-S-V-G-D-R-V-T-I-T-C-R-S-S (SEQ ID NO:78) (or asequence that differs by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2amino acids, e.g., conservative amino acids, therefrom, optionallyprovided that R is not changed); an FR2 comprising the sequenceW-Y-Q-Q-K-P-G-K-A-P-K-L-L-I-Y (SEQ ID NO:79) (or a sequence that differsby no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2 amino acids, e.g.,conservative amino acids, therefrom); an FR3 comprising the sequenceG-V-P-S-R-F-S-G-S-G-S-G-T-D-F-T-L-T-I-S-S-L-Q-P-E-D-F-A-T-Y-Y-C (SEQ IDNO:80) (or a sequence that differs by no more than, 1, 2, 3, 4, or 5,e.g., 1 or 2 amino acids, e.g., conservative amino acids, therefrom,optionally provided that C is not changed, or if changed, is other thanP); and an FR4 comprising the sequence F-G-Q-G-T-K-V-E-I-K (SEQ IDNO:81) (or a sequence that differs by no more than, 1, 2, 3, 4, or 5,e.g., 1 or 2 amino acids, e.g., conservative amino acids, therefrom).

In an embodiment a FR of the light or heavy chain includes one of thehighlighted residues, or one of the highlighted combinations ofresidues, for that FR (e.g., while other residues in that FR might bechanged, the highlighted residue or combination of residues, are notchanged). For example, in an embodiment, one, two or three of I, R, or Lfor heavy chain FR3 is not changed.

In an embodiment, a FR of the light chain and a FR of the heavy chaineach includes one of the highlighted residues, or one of the highlightedcombinations of residues, for that FR. In an embodiment, each of two FRsin the antibody molecule includes one of the highlighted residues, orone of the highlighted combinations of residues, for that FR. In anembodiment, both are in the light chain. In an embodiment, both are inthe heavy chain. In an embodiment, each of FR2 and FR3 in the heavychain includes one of the highlighted residues, or one of thehighlighted combinations of residues, for that FR. In an embodiment,each of FR1 and FR2 in the heavy and light chain includes one of the highlighted residues for that FR. In an embodiment, all of thehighlighted residues in heavy chain FR1-4 are unchanged. In anembodiment, all of the highlighted residues in light chain FR1-4 areunchanged. In an embodiment, all of the highlighted residues in bothheavy and light chain FR1-4 are unchanged.

In another embodiment, the antibody molecule comprises a structural orfunctional property of Ab 031.

In an embodiment, the antibody molecule competes with a referenceantibody molecule, e.g., an antibody molecule described herein, forbinding to a substrate, e.g., an HA. The reference antibody molecule canbe: a) an antibody molecule comprising: i) a heavy chain immunoglobulinvariable region segment comprising a CDR1 comprising the sequenceS-Y-A-M-H (SEQ ID NO: 68); a CDR2 comprising the sequenceV-V-S-Y-D-G-N-Y-K-Y-Y-A-D-S-V-Q-G (SEQ ID NO: 69); and a CDR3 comprisingthe sequence D-S-R-L-R-S-L-L-Y-F-E-W-L-S-Q-G-Y-F-N-P (SEQ ID NO: 70);and ii) a light chain variable region segment comprising: a CDR1comprising the sequence Q-S-I-T-F-N-Y-K-N-Y-L-A (SEQ ID NO:71); a CDR2comprising the sequence W-G-S-Y-L-E-S (SEQ ID NO:72); and a CDR3comprising the sequence Q-Q-H-Y-R-T-P-P-S (SEQ ID NO:73); b) an antibodymolecule comprises one or both of: (i) a heavy chain immunoglobulinvariable region segment comprising SEQ ID NO: 24; and (ii) a light chainvariable region segment comprising SEQ ID NO:45; or c) Ab 031.

In an embodiment, the antibody molecule binds to the same epitope, or aportion thereof, on HA, as does a reference antibody molecule, e.g. anantibody molecule disclosed herein. The reference antibody molecule canbe: a) an antibody molecule comprising: i) a heavy chain immunoglobulinvariable region segment comprising a CDR1 comprising the sequenceS-Y-A-M-H (SEQ ID NO: 68); a CDR2 comprising the sequenceV-V-S-Y-D-G-N-Y-K-Y-Y-A-D-S-V-Q-G (SEQ ID NO: 69); and a CDR3 comprisingthe sequence D-S-R-L-R-S-L-L-Y-F-E-W-L-S-Q-G-Y-F-N-P (SEQ ID NO: 70);and ii) a light chain variable region segment comprising: a CDR1comprising the sequence Q-S-I-T-F-N-Y-K-N-Y-L-A (SEQ ID NO: 71); a CDR2comprising the sequence W-G-S-Y-L-E-S (SEQ ID NO:72); and a CDR3comprising the sequence Q-Q-H-Y-R-T-P-P-S (SEQ ID NO: 73); b) anantibody molecule comprises one or both of: (i) a heavy chainimmunoglobulin variable region segment comprising SEQ ID NO: 24; and(ii) a light chain variable region segment comprising SEQ ID NO: 45; orc) Ab 031.

In an embodiment, the antibody molecule comprises one or both of: aheavy chain variable region comprising least 60, 70, 80, 85, 90, 95, 98or 99 percent homology with SEQ ID NO: 24; and a light chain variableregion comprising least 60, 70, 80, 85, 90, 95, 98 or 99 percenthomology with SEQ ID NO: 45.

In an embodiment, the antibody molecule comprises one or both of: aheavy chain variable region comprising least 60, 70, 80, 85, 90, 95, 98or 99 percent homology with SEQ ID NO: 24; and a light chain variableregion comprising least 60, 70, 80, 85, 90, 95, 98 or 99 percenthomology with SEQ ID NO: 45, wherein, optionally, each HC CDR differs byno more than 1, 2, 3, 4 or 5 amino acids, e.g., 1 or 2, e.g.,conservative amino acids, from the corresponding CDR of SEQ ID NO: 24and each LC CDR differs by no more than 1, 2, 3, 4 or 5 amino acids,e.g., 1 or 2, e.g., conservative amino acids, from the corresponding CDRof SEQ ID NO: 45.

In an embodiment, the antibody molecule comprises one or both of: aheavy chain variable region comprising least 60, 70, 80, 85, 90, 95, 98or 99 percent homology with SEQ ID NO: 25; and a light chain variableregion comprising least 60, 70, 80, 85, 90, 95, 98 or 99 percenthomology with SEQ ID NO: 45, wherein the antibody molecule comprises 1,2, 3, 4, 5, or all of: (i) a HC CDR1 comprising: S at the 1st positionand A at the 3^(rd) position in HC CDR1; (ii) a HC CDR2 comprising oneor both, e.g., one of: V at the 2^(nd) position; or N at the 7^(th)position and Q at the 16^(th) position in HC CDR2; (iii) a HC CDR3comprising: R at the 3^(rd) position (and optionally, L at the 3^(rd)position); (iv) a LC CDR1 comprising: I at the 3^(rd) position; (v) a LCCDR2 comprising one, two, or three of, e.g., one of: G at the 2^(nd)position; Y at the 4^(th) position; or L at the 5^(th) position in LCCDR2; (vi) a LC CDR3 comprising: S at the 9^(th) position in LC CDR3.

In an embodiment, the antibody molecule comprises: (a) a heavy chainimmunoglobulin variable region segment comprising SEQ ID NO: 24 (or asequence that differs by no more than 1, 2, 3, 4 or 5 amino acids, e.g.,conservative amino acids, therefrom); and (b) a light chain variableregion segment comprising SEQ ID NO: 45 (or a sequence that differs byno more than 1, 2, 3, 4 or 5 amino acids, e.g., conservative aminoacids, therefrom).

In an embodiment, the antibody molecule comprises one or both of: (a) aheavy chain immunoglobulin variable region segment comprising SEQ ID NO:24; and (b) a light chain variable region segment comprising SEQ ID NO:45.

In an embodiment, the antibody molecule comprises one or both of: (a) aheavy chain immunoglobulin variable region segment comprising a CDR1comprising the sequence S-Y-A-M-H (SEQ ID NO: 68) (or a sequence thatdiffers by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2 amino acids,e.g., conservative amino acids, therefrom); a CDR2 comprising thesequence V-V-S-Y-D-G-N-Y-K-Y-Y-A-D-S-V-Q-G (SEQ ID NO: 69) (or asequence that differs by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2amino acids, e.g., conservative amino acids, therefrom); and a CDR3comprising the sequence D-S-R-L-R-S-L-L-Y-F-E-W-L-S-Q-G-Y-F-N-P (SEQ IDNO: 70) (or a sequence that differs by no more than, 1, 2, 3, 4, or 5,e.g., 1 or 2 amino acids, e.g., conservative amino acids, therefrom);and (b) a light chain variable region segment comprising a CDR1comprising the sequence Q-S-I-T-F-N-Y-K-N-Y-L-A (SEQ ID NO: 71) (or asequence that differs by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2amino acids, e.g., conservative amino acids, therefrom); a CDR2comprising the sequence W-G-S-Y-L-E-S (SEQ ID NO: 72) (or a sequencethat differs by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2 aminoacids, e.g., conservative amino acids, therefrom); and a CDR3 comprisingthe sequence Q-Q-H-Y-R-T-P-P-S (SEQ ID NO: 73) (or a sequence thatdiffers by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2 amino acids,e.g., conservative amino acids, therefrom).

In an embodiment, the antibody molecule comprises one or both of: a) LCCDR1-3, that collectively, differ from the Ab 031 LC CDR1-3 by no morethan, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, e.g., 1, 2, 3, or 4, aminoacids, e.g., conservative amino acids; and b) HC CDR1-3, thatcollectively, differ from the Ab 031 HC CDR1-3 by no more than, 1, 2, 3,4, 5, 6, 7, 8, 9, or 10, e.g., 1, 2, 3, or 4, amino acids, e.g.,conservative amino acids.

In an embodiment, the antibody molecule comprises one or both of: (a) aheavy chain immunoglobulin variable region segment comprising a CDR1comprising the sequence S-Y-A-M-H (SEQ ID NO: 68) (or a sequence thatdiffers by no more than, 1, 2, or 3, e.g., 1 or 2 amino acids, e.g.,conservative amino acids, therefrom, optionally provided that at least 1or 2 of the highlighted residues are not changed, e.g., both S and A arenot changed); a CDR2 comprising the sequenceV-V-S-Y-D-G-N-Y-K-Y-Y-A-D-S-V-Q-G (SEQ ID NO: 69) (or a sequence thatdiffers by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2 amino acids,e.g., conservative amino acids, therefrom, provided that, e.g., at least1, 2, or 3 of the highlighted residues are not changed, e.g., V or bothN and Q or all three of V, N, and Q are not changed); a CDR3 comprisingthe sequence D-S-R-L-R-S-L-L-Y-F-E-W-L-S-Q-G-Y-F-N-P (SEQ ID NO: 70) (ora sequence that differs by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2amino acids, e.g., conservative amino acids, therefrom optionallyprovided that, e.g., R is not changed); and (b) a light chain variableregion segment comprising a CDR1 comprising the sequenceQ-S-I-T-F-N-Y-K-N-Y-L-A (SEQ ID NO: 71) or a sequence that differs by nomore than, 1, 2, 3, 4, or 5, e.g., 1 or 2 amino acids, e.g.,conservative amino acids, therefrom, optionally provided that at least 1or 2 of the highlighted residues are not changed, e.g., I is notchanged); a CDR2 comprising the sequence W-G-S-Y-L-E-S (SEQ ID NO:72)(or a sequence that differs by no more than, 1, 2, 3, 4, or 5, e.g., 1or 2 amino acids, e.g., conservative amino acids, therefrom, optionallyprovided that at least 1, 2, or 3 of the highlighted residues are notchanged, e.g., 1, 2 or all of G, Y, and L are not changed); a CDR3comprising the sequence Q-Q-H-Y-R-T-P-P-S (SEQ ID NO:73) (or a sequencethat differs by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2 aminoacids, e.g., conservative amino acids, therefrom, optionally providedthat at least one or both of the highlighted residues are not changed,e.g., S is not changed).

In an embodiment, a CDR of the light or heavy chain includes one of thehighlighted residues, or one of the highlighted combinations ofresidues, for that CDR (e.g., while other residues in that CDR might bechanged, the highlighted residue or combination of residues, are notchanged). In an embodiment, a CDR of the light and a CDR of the heavychain each includes one of the highlighted residues, or one of thehighlighted combinations of residues, for that CDR. In an embodiment,each of two CDRs in the antibody molecule includes one of thehighlighted residues, or one of the highlighted combinations ofresidues, for that CDR. In an embodiment, both are in the light chain.In an embodiment, both are in the heavy chain. In an embodiment, each ofthe three CDRs in the heavy chain includes one of the highlightedresidues, or one of the highlighted combinations of residues, for thatCDR. In an embodiment, each of the three CDRs in the light chainincludes one of the highlighted residues, or one of the highlightedcombinations of residues, for that CDR. In an embodiment, each of thesix CDRs in the heavy and light chain includes one of the highlightedresidues, or one of the highlighted combinations of residues, for thatCDR.

In an embodiment, the antibody molecule comprises one or more (e.g., 2,3, 4, 5, or all) of the following properties: (a) both S and A in HCCDR1 are unchanged; (b) V or both N and Q or all three of V, N, and Q inHC CDR2 are unchanged; (c) R in HC CDR3 is unchanged; (d) I in LC CDR1is unchanged; (e) 1, 2 or 3 of G, Y, and L in LC CDR2 are unchanged; (f)S in LC CDR3 is unchanged.

In an embodiment, the antibody molecule comprises 1, 2, 3, 4, 5, or all6 properties selected from (a) to (f). In an embodiment, the antibodymolecule comprises a heavy chain having a one or more propertiesselected from (a), (b), and (c) and a light chain having one or moreproperties selected from (d), (e), and (f). In the embodiment, theantibody molecule comprises one or both of: (a) a heavy chainimmunoglobulin variable region segment comprising a CDR1 comprising thesequence S-Y-A-M-H (SEQ ID NO: 68); a CDR2 comprising the sequenceV-V-S-Y-D-G-N-Y-K-Y-Y-A-D-S-V-Q-G (SEQ ID NO: 69); and a CDR3 comprisingthe sequence D-S-R-L-R-S-L-L-Y-F-E-W-L-S-Q-G-Y-F-N-P (SEQ ID NO:70); and(b) a light chain variable region segment comprising a CDR1 comprisingthe sequence Q-S-I-T-F-N-Y-K-N-Y-L-A (SEQ ID NO: 71); a CDR2 comprisingthe sequence W-G-S-Y-L-E-S (SEQ ID NO: 72); and a CDR3 comprising thesequence Q-Q-H-Y-R-T-P-P-S (SEQ ID NO: 73).

In an embodiment, the antibody molecule comprises one or both of: a) oneor more framework regions (FRs) from SEQ ID NO: 24. For example, theantibody molecule comprises one or more or all of FR1, FR2, FR3, or FR4,or sequences that differ individually, or collectively, by no more than1, 2, 3, 4, of 5 amino acid residues, e.g., conservative residues, fromSEQ ID NO: 24; and b) one or more framework regions (FRs) from SEQ IDNO: 45. For example, the antibody molecule comprises one or more or allof FR1, FR2, FR3, or FR4, or sequences that differ individually, orcollectively, by no more than 1, 2, 3, 4, of 5 amino acid residues,e.g., conservative residues, from SEQ ID NO: 45.

In an embodiment, the antibody molecule comprises: (a) a heavy chainimmunoglobulin variable region segment that further comprises one ormore or all of: an FR1 comprising the sequenceE-V-Q-L-L-E-S-G-G-G-L-V-K-P-G-Q-S-L-K-L-S-C-A-A-S-G-F-T-F-T (SEQ IDNO:82) (or a sequence that differs by no more than, 1, 2, 3, 4, or 5,e.g., 1 or 2 amino acids, e.g., conservative amino acids, therefrom,optionally provided that T is not changed); an FR2 comprising thesequence W-V-R-Q-P-P-G-K-G-L-E-W-V-A (SEQ ID NO:75) (or a sequence thatdiffers by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2 amino acids,e.g., conservative amino acids, therefrom, optionally provided that W isnot changed, or that if changed, is other than R); an FR3 comprising thesequence R-F-T-I-S-R-D-N-S-K-N-T-L-Y-L-Q-M-N-S-L-R-A-E-D-T-A-V-Y-Y-C-A-K(SEQ ID NO:76) (or a sequence that differs by no more than, 1, 2, 3, 4,or 5, e.g., 1 or 2 amino acids, e.g., conservative amino acids,therefrom, optionally provided that one, two or three of I, R, or L isnot changed, or that if I is changed it is other than G, if R is changedit is other than P. or if L is changed it is other than A); and an FR4comprising the sequence W-G-Q-G-T-T-L-T-V-S-S (SEQ ID NO:77) (or asequence that differs by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2amino acids, e.g., conservative amino acids, therefrom) orW-G-Q-G-T-T-V-T-V-S-S (SEQ ID NO:171) (or a sequence that differs by nomore than, 1, 2, 3, 4, or 5, e.g., 1 or 2 amino acids, e.g.,conservative amino acids, therefrom); and (a) a light chainimmunoglobulin variable region segment further comprises one or more orall of: an FR1 comprising the sequenceD-I-Q-M-T-Q-S-P-S-S-L-S-A-S-V-G-D-R-V-T-I-T-C-R-S-S (SEQ ID NO:78) (or asequence that differs by no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2amino acids, e.g., conservative amino acids, therefrom, optionallyprovided that R is not changed); an FR2 comprising the sequenceW-Y-Q-Q-K-P-G-K-A-P-K-L-L-I-Y (SEQ ID NO:79) (or a sequence that differsby no more than, 1, 2, 3, 4, or 5, e.g., 1 or 2 amino acids, e.g.,conservative amino acids, therefrom); an FR3 comprising the sequenceG-V-P-S-R-F-S-G-S-G-S-G-T-D-F-T-L-T-I-S-S-L-Q-P-E-D-F-A-T-Y-Y-C (SEQ IDNO:80) (or a sequence that differs by no more than, 1, 2, 3, 4, or 5,e.g., 1 or 2 amino acids, e.g., conservative amino acids, therefrom,optionally provided that C is not changed, or if changed, is other thanP); and an FR4 comprising the sequence F-G-Q-G-T-K-V-E-I-K (SEQ IDNO:81) (or a sequence that differs by no more than, 1, 2, 3, 4, or 5,e.g., 1 or 2 amino acids, e.g., conservative amino acids, therefrom).

In an embodiment, a FR of the light or heavy chain includes one of thehighlighted residues, or one of the highlighted combinations ofresidues, for that FR (e.g., while other residues in that FR might bechanged, the highlighted residue or combination of residues, are notchanged). For example, in an embodiment, one, two or three of I, R, or Lfor heavy chain FR3 is not changed.

In an embodiment, a FR of the light chain and a FR of the heavy chaineach includes one of the highlighted residues, or one of the highlightedcombinations of residues, for that FR. In an embodiment, each of two FRsin the antibody molecule includes one of the highlighted residues, orone of the highlighted combinations of residues, for that FR. In anembodiment, both are in the light chain. In an embodiment, both are inthe heavy chain. In an embodiment each of FR2 and FR3 in the heavy chainincludes one of the highlighted residues, or one of the highlightedcombinations of residues, for that FR. In an embodiment, each of FR1 andFR2 in the heavy and light chain includes one of the highlightedresidues for that FR. In an embodiment, all of the highlighted residuesin heavy chain FR1-4 are unchanged. In an embodiment, all of thehighlighted residues in light chain FR1-4 are unchanged. In anembodiment, all of the highlighted residues in both heavy and lightchain FR1-4 are unchanged.

In an embodiment, the antibody molecule comprises: (a) the heavy chainimmunoglobulin variable region segment comprises one or more or all ofan FR1 comprising the sequenceE-V-Q-L-L-E-S-G-G-G-L-V-K-P-G-Q-S-L-K-L-S-C-A-A-S-G-F-T-F-T (SEQ ID NO:82); an FR2 comprising the sequence W-V-R-Q-P-P-G-K-G-L-E-W-V-A (SEQ IDNO: 75); an FR3 comprising the sequenceR-F-T-I-S-R-D-N-S-K-N-T-L-Y-L-Q-M-N-S-L-R-A-E-D-T-A-V-Y-Y-C-A-K (SEQ IDNO: 76); and an FR4 comprising the sequence W-G-Q-G-T-T-L-T-V-S-S (SEQID NO: 77) or W-G-Q-G-T-T-V-T-V-S-S (SEQ ID NO: 171); and (b) the lightchain immunoglobulin variable region segment comprising one or more orall of an FR1 comprising the sequenceD-I-Q-M-T-Q-S-P-S-S-L-S-A-S-V-G-D-R-V-T-I-T-C-R-S-S (SEQ ID NO: 78); anFR2 comprising the sequence W-Y-Q-Q-K-P-G-K-A-P-K-L-L-I-Y (SEQ ID NO:79); an FR3 comprising the sequenceG-V-P-S-R-F-S-G-S-G-S-G-T-D-F-T-L-T-I-S-S-L-Q-P-E-D-F-A-T-Y-Y-C (SEQ IDNO: 80); and an FR4 comprising the sequence F-G-Q-G-T-K-V-E-I-K (SEQ IDNO: 81).

In another embodiment, the antibody molecule comprises: (a) a heavychain immunoglobulin variable region segment comprising SEQ ID NO: 24(or a sequence that differs by no more than 1, 2, 3, 4 or 5 amino acids,e.g., conservative amino acids, therefrom); and (b) a light chainvariable region segment comprising SEQ ID NO:45 (or a sequence thatdiffers by no more than 1, 2, 3, 4 or 5 amino acids, e.g., conservativeamino acids, therefrom).

In another embodiment, the antibody molecule comprises a structural orfunctional property of one or both a heavy chain variable region and alight chain variable region disclosed herein.

In an embodiment, the antibody molecule competes with a referenceantibody molecule, e.g., an antibody molecule described herein, forbinding to a substrate, e.g., an HA. The reference antibody molecule canbe: a) an antibody molecule comprising the heavy and light CDRs from: aheavy chain variable region from Table 3, Table 4A, Table 4B, FIG. 2,FIG. 5, or FIG. 7; and a light chain variable region from Table 3, Table4A, Table 4B, FIGS. 3A-3B, FIGS. 6A-6B, or FIG. 7; b) an antibodymolecule that comprises: (i) a heavy chain immunoglobulin variableregion segment from Table 3, Table 4A, Table 4B, FIG. 2, FIG. 5, or FIG.7; and (ii) a light chain variable region segment from Table 3, Table4A, Table 4B, FIGS. 3A-3B, FIGS. 6A-6B, or FIG. 7; or c) an antibodydisclosed herein.

In an embodiment the antibody molecule binds to the same epitope, or aportion thereof, on HA, as does a reference antibody molecule, e.g. anantibody molecule disclosed herein. The reference antibody molecule canbe: a) an antibody molecule comprising the heavy and light CDRs from: aheavy chain variable region from Table 3, Table 4A, Table 4B, FIG. 2,FIG. 5, or FIG. 7; and a light chain variable region from Table 3, Table4A, Table 4B, FIGS. 3A-3B, FIGS. 6A-6B, or FIG. 7; b) an antibodymolecule that comprises: (i) a heavy chain immunoglobulin variableregion segment from Table 3, Table 4A, Table 4B, FIG. 2, FIG. 5, or FIG.7; and (ii) a light chain variable region segment from Table 3, Table4A, Table 4B, FIGS. 3A-3B, FIGS. 6A-6B, or FIG. 7; or c) an antibodydisclosed herein.

In an embodiment, the antibody molecule comprises one or both of: aheavy chain variable region comprising least 60, 70, 80, 85, 90, 95, 98or 99 percent homology with a reference heavy chain from Table 3, Table4A, Table 4B, FIG. 2, FIG. 5 or FIG. 7; and a light chain variableregion comprising least 60, 70, 80, 85, 90, 95, 98 or 99 percenthomology with reference light chain from Table 3, Table 4A, Table 4B,FIGS. 3A-3B, FIGS. 6A-6B or FIG. 7, wherein, optionally, each HC CDRdiffers by no more than 1, 2, 3, 4 or 5 amino acids, e.g., 1 or 2, e.g.,conservative amino acids, from the corresponding HC CDR from itsreference heavy chain and each LC CDR differs by no more than 1, 2, 3, 4or 5 amino acids, e.g., 1 or 2, e.g., conservative amino acids, from thecorresponding CDR in its reference light chain.

In an embodiment, the antibody molecule comprises: a heavy chainvariable region comprising least 60, 70, 80, 85, 90, 95, 98 or 99percent homology with a heavy chain from Table 3 and a light chainvariable region comprising least 60, 70, 80, 85, 90, 95, 98 or 99percent homology with the corresponding light chain from Table 3.

In an embodiment, the antibody molecule comprises: a heavy chainvariable region comprising least 60, 70, 80, 85, 90, 95, 98 or 99percent homology with a heavy chain from Table 4A and a light chainvariable region comprising least 60, 70, 80, 85, 90, 95, 98 or 99percent homology with the corresponding light chain from Table 4A.

In an embodiment the antibody molecule comprises: a heavy chain variableregion comprising least 60, 70, 80, 85, 90, 95, 98 or 99 percenthomology with a heavy chain from Table 4B and a light chain variableregion comprising least 60, 70, 80, 85, 90, 95, 98 or 99 percenthomology with the corresponding light chain from Table 4B.

In an embodiment the antibody molecule comprises one or both of: a heavychain variable region from Table 3, Table 4A, Table 4B, FIG. 2, FIG. 5,or FIG. 7; and a light chain variable region from Table 3, Table 4A,Table 4B, FIGS. 3A-3B, FIGS. 6A-6B, or FIG. 7.

In an embodiment the antibody molecule comprises: a heavy chain variableregion from Table 3 and the corresponding light chain from Table 3; aheavy chain from Table 4A and the corresponding light chain from Table4A; or a heavy chain from Table 4B and the corresponding light chainfrom Table 4B.

In an embodiment, the antibody molecule comprises one or both of: (a) aheavy chain immunoglobulin variable region segment comprising a CDR1, aCDR2 and a CDR3 from a heavy chain sequence of Table 3, Table 4A, Table4B, FIG. 2, FIG. 5, or FIG. 7 (or CDRs that, individually orcollectively, differ therefrom by no more than, 1, 2, 3, 4, or 5, e.g.,1 or 2 amino acids, e.g., conservative amino acids)); and (b) a lightchain immunoglobulin variable region segment comprising a CDR1, a CDR2and a CDR3 from a light chain sequence of Table 3, Table 4A, Table 4B,FIGS. 3A-3B, FIGS. 6A-6B, or FIG. 7 (or CDRs that, individually orcollectively, differ therefrom by no more than, 1, 2, 3, 4, or 5, e.g.,1 or 2 amino acids, e.g., conservative amino acids).

In an embodiment, the antibody molecule comprises one or both of: CDRsfrom a heavy chain of Table 3 and the light chain CDRs from thecorresponding light chain from Table 3. In an embodiment, the antibodymolecule comprises one or both of: CDRs from a heavy chain of Table 4Aand the light chain CDRs from the corresponding light chain from Table4A. In an embodiment, the antibody molecule comprises one or both of:CDRs from a heavy chain of Table 4B and the light chain CDRs from thecorresponding light chain from Table 4B.

In an embodiment, the antibody molecule comprises one or both of: (a) aheavy chain immunoglobulin variable region segment comprising a CDR1, aCDR2; and a CDR3 from a heavy chain sequence of FIG. 2, FIG. 5, or FIG.7; and (b) a light chain immunoglobulin variable region segmentcomprising a CDR1, a CDR2 and a CDR3 from a light chain sequence ofFIGS. 3A-3B, FIGS. 6A-6B, or FIG. 7. In an embodiment, the antibodymolecule comprises: (a) a heavy chain immunoglobulin variable regionsegment from FIG. 2 or FIG. 7; and (b) a light chain immunoglobulinvariable region segment from FIGS. 3A-3B or FIG. 7.

In an embodiment, the heavy chain immunoglobulin variable region furthercomprises an Isoleucine-Aspartate (Ile-Asp) dipeptide at the N-terminus.In another embodiment, the light chain immunoglobulin variable regionfurther comprises an Ile-Asp dipeptide at the N-terminus. In yet anotherembodiment, both the heavy chain immunoglobulin variable region and thelight chain immunoglobulin variable region or an antibody featured inthe disclosure further comprises an Ile-Asp dipeptide at the N-terminus.In other embodiment the Ile-Asp dipeptide is absent from one or both theheavy and light chain.

In an embodiment, the antibody molecule comprises one or both of: a) oneor more framework regions (FRs) from heavy chain disclosed herein. E.g.,the antibody molecule comprises one or more or all of FR1, FR2, FR3, orFR4, or sequences that differ individually, or collectively, by no morethan 1, 2, 3, 4, of 5 amino acid residues, e.g., conservative residues,from heavy chain disclosed herein; and b) one or more framework regions(FRs) from light chain disclosed herein. E.g., the antibody moleculecomprises one or more or all of FR1, FR2, FR3, or FR4, or sequences thatdiffer individually, or collectively, by no more than 1, 2, 3, 4, of 5amino acid residues, e.g., conservative residues, from light chaindisclosed herein.

In an embodiment, the antibody molecule comprises:

(a) a heavy chain immunoglobulin variable region segment comprising oneor more or all of a CDR1 comprising the sequenceG-F-T-F-[S/T]-[S/T]-Y-[A/G]-M-H (SEQ ID NO: 184), or a sequence thatdiffers from SEQ ID NO: 184 by no more than 1 or 2 residues; a CDR2comprising the sequenceV-[I/V/L]-S-[Y/F]-D-G-[S/N]-[Y/N]-[K/R]-Y-Y-A-D-S-V-Q-G (SEQ ID NO: 2)or a sequence that differs from SEQ ID NO: 2 by no more than 1 or 2residues; or a CDR3 comprising the sequenceD-[S/T]-[R/K/Q]-L-R-[S/T]-L-L-Y-F-E-W-L-S-[Q/S]-G-[Y/L/V]-[F/L]-[N/D]-[P/Y](SEQ ID NO: 3), or a sequence that differs from SEQ ID NO:3 by no morethan 1 or 2 residues; and

(b) a light chain variable region segment comprising one or more or allof a CDR1 comprising the sequence[K/R]-S-S-Q-[S/T]-[V/L/I]-[T/S]-[Y/F/W]-[N/S/D]-Y-K-N-Y-L-A (SEQ ID NO:185) or a sequence that differs from SEQ ID NO: 185 by no more than 1 or2 residues, or comprising the sequence[K/R]-S-S-Q-[S/T]-[V/L/I]-[T/S]-[Y/F/W]-[N/S/D/Q/R/E]-Y-K-N-Y-L-A (SEQID NO: 186) or a sequence that differs from SEQ ID NO: 186 by no morethan 1 or 2 residues or[K/R]-S-S-Q-[S/T]-[V/L/I]-[T/S]-[Y/F/w]-[N/S/D/E]-Y-K-N-Y-L-A (SEQ IDNO: 185) or a sequence that differs from SEQ ID NO: 186 by no more than1 or 2 residues; a CDR2 comprising the sequenceW-[A/G]-S-[T/A/Y/H/K/D]-[R/L]-E-[S/T] (SEQ ID NO: 5) or a sequence thatdiffers from SEQ ID NO:5 by no more than 1 or 2 residues; or a CDR3comprising the sequence Q-Q-[Y/H]-Y-R-T-P-P-[T/S] (SEQ ID NO: 6) or asequence that differs from SEQ ID NO:6 by no more than 1 or 2 residues;

optionally, provided that,

if the light chain variable region segment comprises: a CDR 1 comprisingthe sequence K-S-S-Q-S-V-T-Y-N-Y-K-N-Y-L-A (SEQ ID NO:83); a CDR2comprising the sequence W-A-S-T-R-E-S (SEQ ID NO: 84); and a CDR3comprising the sequence Q-Q-Y-Y-R-T-P-P-T (SEQ ID NO: 85);

then the heavy chain variable region segment comprises one or more ofthe following: (a) CDRs other than the following: a CDR1 comprising thesequence S-Y-G-M-H (SEQ ID NO: 86); a CDR2 comprising the sequenceV-I-S-Y-D-G-S-Y-K-Y-Y-A-D-S-V-Q-G (SEQ ID NO: 87); or a CDR3 comprisingthe sequence D-S-E-L-R-S-L-L-Y-F-E-W-L-S-Q-G-Y-F-N-P (SEQ ID NO:88); or(b) FRs other than the following: an FR1 other thanE-V-Q-L-L-E-S-G-G-G-L-V-K-P-G-Q-S-L-K-L-S-C-A-A-S-G-F-T-F-T (SEQ ID NO:82); an FR2 other than W-V-R-Q-P-P-G-K-G-L-E-W-V-A (SEQ ID NO:75); anFR3 other thanR-F-T-I-S-R-D-N-S-K-N-T-L-Y-L-Q-M-N-S-L-R-A-E-D-T-A-V-Y-Y-C-A-K (SEQ IDNO: 76); or an FR4 other than W-G-A-G-T-T-L-T-V-S-S (SEQ ID NO: 89); (c)a CDR1 where the amino residue at position 5 of SEQ ID NO: 184 is an S,the amino acid residue at position 6 of SEQ ID NO: 184 is a T, or theamino acid residue at position 8 of SEQ ID NO: 184 is an A; (d) a CDR2wherein the amino residue at position 2 of SEQ ID NO: 2 is a V or an L,the amino acid at position 4 is an F, the amino acid at position 7 is anN, the amino acid at position 8 is a Y, or the amino acid at position 9is a R; (e) a CDR3 wherein the amino residue at position 2 of SEQ IDNO:3 is a T, the amino acid residue at position 3 of SEQ ID NO:3 is anR, a K, or a Q, the amino acid residue at position 6 of SEQ ID NO: 3 isa T, the amino acid residue at position 15 of SEQ ID NO: 3 is an S, theamino acid residue at position 17 of SEQ ID NO:3 is an L, or a V, theamino acid residue at position 18 of SEQ ID NO:3 is an L, the amino acidresidue at position 19 of SEQ ID NO:3 is a D, or the amino acid residueat position 20 of SEQ ID NO:3 is a Y; (f) an FR1 wherein the aminoresidue at position 11 of SEQ ID NO: 7 is a Q, or the amino acid residueat position 7 of SEQ ID NO: 7 is a T; (g) an FR4 wherein the aminoresidue at position 3 of SEQ ID NO:10 is a Q, the amino acid residue atposition 5 of SEQ ID NO: 10 is an A; the amino acid residue at position6 of SEQ ID NO: 10 is an M, or the amino acid residue at position 7 ofSEQ ID NO:10 is a V; or (h) it produces fewer escape mutants than does areference anti-HA antibody molecule, e.g., Ab 67-11, FI6, FI28, C179,F10, CR9114, or CR6261, e.g., when tested by a method disclosed herein,and also provided that, if the heavy chain immunoglobulin variableregion segment comprises: a CDR1 comprising the sequence S-Y-G-M-H (SEQID NO: 86); a CDR2 comprising the sequenceV-I-S-Y-D-G-S-Y-K-Y-Y-A-D-S-V-Q-G (SEQ ID NO:87); and a CDR3 comprisingthe sequence D-S-E-L-R-S-L-L-Y-F-E-W-L-S-Q-G-Y-F-N-P (SEQ ID NO: 88),then the light chain variable region segment comprises one of more ofthe following: (a) CDRs other than the following: CDR1 KSSQSVTYNYKNYLA(SEQ ID NO: 83); CDR2 WASTRES (SEQ ID NO:84); or CDR3 QQYYRTPPT (SEQ IDNO: 85); (b) FRs other than the following: FR1 comprising the sequenceEIVMTQSPDSLAVSLGERATINC (SEQ ID NO: 90); FR2 comprising the sequenceWYQQKPGQPPKLLIY (SEQ ID NO: 91); FR3 comprising the sequenceGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYC (SEQ ID NO: 92); or FR4 comprising thesequence FGGGTKLDIK (SEQ ID NO: 93); (c) a CDR1 wherein the aminoresidue at position 1 of SEQ ID NO: 185 is an R, the amino residue atposition 5 of SEQ ID NO:4 is a T, the amino residue at position 6 of SEQID NO:4 is an L or an I, the amino residue at position 7 of SEQ ID NO:185 is an S, the amino residue at position 8 of SEQ ID NO: 185 is an For a W, or the amino residue at position 9 of SEQ ID NO: 185 is an S ora D; (d) a CDR2 wherein the amino residue at position 2 of SEQ ID NO: 5is a G, the amino residue at position 4 of SEQ ID NO: 5 is an A, a Y, anH, a K, or a D, the amino residue at position 5 of SEQ ID NO: 5 is an L,the amino residue at position 7 of SEQ ID NO: 5 is a T; (e) a CDR3wherein the amino residue at position 3 of SEQ ID NO: 6 is an H; theamino acid residue at position 9 of SEQ ID NO: 6 is an S; (f) an FR1wherein the amino residue at position 1 of SEQ ID NO: 11 is a D; theamino residue at position 3 of SEQ ID NO: 11 is a Q, the amino residueat position 9 of SEQ ID NO: 11 is an S, the amino residue at position 10of SEQ ID NO: 11 is a T, the amino residue at position 11 of SEQ ID NO:11 is a V, the amino residue at position 12 of SEQ ID NO:11 is an S, theamino residue at position 13 of SEQ ID NO: 11 is an A, the amino residueat position 14 of SEQ ID NO:11 is a T, the amino residue at position 15of SEQ ID NO:11 is a V or an R, the amino residue at position 17 of SEQID NO: 11 is a D, the amino residue at position 20 of SEQ ID NO:11 is anS, the amino residue at position 22 of SEQ ID NO:11 is a T, a Q, a D, oran R; (g) an FR2 wherein the amino residue at position 8 of SEQ ID NO:12is a K; or the amino residue at position 9 of SEQ ID NO: 12 is an A; (h)an FR3 wherein the amino residue at position 4 of SEQ ID NO: 13 is an Eor an S; the amino residue at position 24 of SEQ ID NO: 13 is a P, theamino residue at position 27 of SEQ ID NO: 13 is an F, a K, or a D, theamino residue at position 29 of SEQ ID NO: 13 is a T; (i) an FR4 whereinthe amino residue at position 3 of SEQ ID NO:14 is a Q, a T, an S, or anN, the amino residue at position 7 of SEQ ID NO:14 is a V, or the aminoresidue at position 8 of SEQ ID NO:14 is an E; or (j) it produces fewerescape mutants than does a reference anti-HA antibody molecule, e.g., Ab67-11, FI6, FI28, C179, F10, CR9114, or CR6261, e.g., when tested by amethod disclosed herein; and further provided that if the light chainvariable region segment comprises: a CDR 1 comprising the sequenceK-S-S-Q-S-V-T-F-N-Y-K-N-Y-L-A (SEQ ID NO: 146); a CDR2 comprising thesequence W-A-S-A-R-E-S (SEQ ID NO: 147); and a CDR3 comprising thesequence Q-Q-H-Y-R-T-P-P-T (SEQ ID NO: 148); then the heavy chainvariable region segment comprises one or more of the following: CDRsother than the CDR's described at FIG. 4; or FRs other than the FRsdescribed at FIG. 4.

In an embodiment, the heavy chain CDR sequences, collectively, differfrom the recited sequences by no more than 5, 4, 3, 2 or 1 amino acidresidues; and the light chain CDR sequences, collectively, differ fromthe recited sequences by no more than 5, 4, 3, 2 or 1 amino acidresidues.

In an embodiment, the antibody molecule comprises:

(a) a heavy chain (HC) immunoglobulin variable region segmentcomprising:

-   -   an HC CDR1 comprising the sequence S-Y-A-M-H (SEQ ID NO:68), or        a sequence that differs therefrom at the 3^(rd) position (A to G        substitution);    -   an HC CDR2 comprising the sequence        V-V-S-Y-D-G-N-Y-K-Y-Y-A-D-S-V-Q-G (SEQ ID NO:69) or a sequence        that differs therefrom at the 2^(nd) position (V to I        substitution), the 7^(th) residue (N to S substitution), the        8^(th) position (Y to N substitution), or a combination thereof;    -   an HC CDR3 comprising the sequence        D-S-R-L-R-S-L-L-Y-F-E-W-L-S-Q-G-Y-F-N-P (SEQ ID NO:70), or a        sequence that differs therefrom at the 2^(nd) position (S to T        substitution), the 3^(rd) position (R to K substitution), the        15^(th) position (Q to S substitution), the 17^(th) position (Y        to L substitution), the 18^(th) position (F to L substitution),        the 19^(th) position (N to D substitution), the 20^(th) position        (P to Y substitution), or a combination thereof; and

(b) a light chain (LC) immunoglobulin variable region segmentcomprising:

-   -   an LC CDR1 comprising the sequence Q-S-I-T-F-N-Y-K-N-Y-L-A (SEQ        ID NO:71), or a sequence that differs therefrom at the 2^(nd)        position (S to T substitution), the 3^(rd) position (I to V        substitution), the 5^(th) position (F to Y substitution), the        6^(th) position (N to S or N to D substitution), the 12^(th)        position (A to G substitution), or a combination thereof;    -   an LC CDR2 comprising the sequence W-G-S-Y-L-E-S (SEQ ID NO:72),        or a sequence that differs therefrom at the 2^(nd) position (G        to A substitution), the 4^(th) position (Y to T substitution),        the 5^(th) position (L to R substitution), or a combination        thereof;    -   an LC CDR3 comprising the sequence Q-Q-H-Y-R-T-P-P-S (SEQ ID        NO:73), or a sequence that differs therefrom at the 3^(rd)        position (H to Y substitution), the 9^(th) position (S to T        substitution), or both.

In an embodiment, the HC CDR1-3 and LC CDR1-3, collectively, comprisesequences that differ by 0, 1 or 2 amino acids from SEQ ID NOS: 68-73.

In an embodiment, the antibody molecule comprises a heavy chainimmunoglobulin variable region segment encoded by a nucleotide sequencedescribed herein. In another embodiment, the antibody molecule comprisesa light chain immunoglobulin variable region segment encoded by anucleotide sequence described herein. In yet another embodiment, theantibody molecule comprises a heavy chain immunoglobulin variable regionsegment encoded by a nucleotide sequence described herein and a lightchain immunoglobulin variable region segment encoded by a nucleotidesequence described herein.

In an embodiment, the heavy chain immunoglobulin variable region segmentis expressed from a recombinant vector, such as an expression vector,that comprises a nucleotide sequence that encodes a heavy chainimmunoglobulin variable region segment. In another embodiment, the lightchain immunoglobulin variable segment is expressed from a recombinantvector, such as an expression vector, that comprises a nucleotidesequence that encodes a light chain immunoglobulin variable regionsegment. In yet another embodiment, the heavy chain immunoglobulinvariable region segment and light chain immunoglobulin variable regionsegment are expressed from a recombinant vector, such as an expressionvector, that comprises a nucleotide sequence that encodes a heavy chainimmunoglobulin variable region segment and a nucleotide sequence thatencodes a light chain immunoglobulin variable region segment.

In an embodiment, the nucleotide sequence encodes (a) a heavy chainimmunoglobulin variable region segment comprising the amino acidsequence of: S-Y-A-M-H (SEQ ID NO: 68) in CDR1;V-V-S-Y-D-G-N-Y-K-Y-Y-A-D-S-V-Q-G (SEQ ID NO: 69) in CDR2; andD-S-R-L-R-S-L-L-Y-F-E-W-L-S-Q-G-Y-F-N-P (SEQ ID NO: 70) in CDR3; and (b)a light chain immunoglobulin variable region segment comprising theamino acid sequence of: Q-S-I-T-F-D-Y-K-N-Y-L-A (SEQ ID NO: 145) inCDR1; W-G-S-Y-L-E-S (SEQ ID NO: 72) in CDR2; and Q-Q-H-Y-R-T-P-P-S (SEQID NO: 73) in CDR3.

In an embodiment, the heavy chain immunoglobulin variable region segmentis expressed from a cell (e.g., a host cell) containing a recombinantvector described herein, such as a recombinant vector comprising anucleic acid sequence that encodes a heavy chain immunoglobulin variableregion. In another embodiment, the light chain immunoglobulin variableregion segment is expressed from a cell (e.g., a host cell) containing arecombinant vector described herein, such as a recombinant vectorcomprising a nucleic acid sequence that encodes a light chainimmunoglobulin variable region. In yet another embodiment, the cell(e.g., a host cell) contains a recombinant vector comprising a nucleicacid sequence that encodes a heavy chain immunoglobulin variable region,and a nucleic acid sequence that encodes a light chain immunoglobulinvariable region.

In an embodiment, the antibody molecule is made by a method comprisingproviding a cell (e.g., a host cell) comprising a nucleotide sequenceexpressing a heavy chain variable region segment and a nucleotidesequence expressing a light chain variable region segment, andexpressing the nucleic acids in the cell.

In an embodiment, the nucleotide sequence expressing the heavy chainvariable region segment and the nucleotide sequence expressing the lightchain variable region segment are on the same recombinant vector (e.g.,expression vector). In another embodiment, the nucleotide sequenceexpressing the heavy chain variable region segment and the nucleotidesequence expressing the light chain variable region segment are onseparate recombinant vectors (e.g., expression vectors).

In an embodiment, the antibody molecule is present in a pharmaceuticalcomposition containing a pharmaceutically acceptable carrier. In anembodiment, the pharmaceutical composition is present in a container asdescribed herein.

Methods of Use

In another aspect, the disclosure features a method of treating orpreventing infection with an influenza virus (e.g., an influenza Avirus, e.g., a Group 1 strain, e.g., an H1N1 strain, e.g., A/SouthCarolina/1/1918, A/Puerto Rico/08/1934, or A/California/04/2009, or anH5N1 strain, e.g., A/Indonesia/5/2005 or A/Vietnam/1203/2004, or aninfluenza B virus, e.g., B/Wisconsin/1/2010), in a subject, e.g., ahuman subject. The method includes administering a formulation describedherein, e.g., a pharmaceutical formulation described herein, to asubject, e.g., human subject, in need thereof.

In an embodiment, the influenza A virus is an H1, H5, H9, H3 or H7strain, such as an H1N1 strain, an H3N2 strain, an H5N1 strain, or anH7N9 strain of influenza A virus.

In an embodiment, the formulation is administered at dose of about 2000mg to about 5000 mg, e.g., about 2300 mg to about 4600 mg, about 2000 mgto about 2500 mg, about 2500 mg to about 3000 mg, about 3000 mg to about3500 mg, about 3500 mg to about 4000 mg, about 4000 mg to about 4500 mg,about 4500 to about 5000 mg, of the antibody molecule. In an embodiment,the formulation is administered at a dose about 2300 mg or about 4600mg, of the antibody molecule. In an embodiment, the formulation isadministered intravenously, e.g., by infusion.

In an embodiment, the administration results in, or correlates with, oneor more of a reduction in the incidence or severity of a symptom ormanifestation of an influenza infection, or the delay or onset of asymptom or manifestation of an influenza infection. In an embodiment,the administration results in, or correlates with, one or more of areduction in the incidence or severity of a symptom or manifestation ofa secondary infection, or the delay or onset of a symptom ormanifestation of a secondary infection.

In an embodiment, the subject, e.g., a human subject, has beenadministered, or the method comprises, administering, or recommendingthe administration of, a second or additional therapy. In an embodiment,the antibody molecule is administered in combination with a second oradditional agent or therapy.

In an embodiment, the second or additional therapy comprisesadministration of a vaccine or an anti-viral therapy, e.g., an anti-NAor an anti-M2 therapy. In an embodiment the second or additional therapycomprises a administration of a vaccine, e.g., a vaccine describedherein or a mixture (a.k.a. a cocktail) of influenza peptides tostimulate the patient's immune system to prevent infection withparticular strains of influenza A. In an embodiment the second oradditional agent comprises administering an anti-viral agent, a painreliever, an anti-inflammatory, an antibiotic, a steroidal agent, asecond therapeutic antibody molecule (e.g., an anti-HA antibody), anadjuvant, a protease or glycosidase (e.g., sialidase). In an embodimentthe second or additional agent comprises, acyclovir, ribavirin,amantadine, remantidine, a neuraminidase inhibitor (e.g., zanamivir(Relenza®), oseltamivir (Tamiflu®), laninamivir, peramivir), orrimantadine.

In an embodiment the second or additional agent comprises a secondantibody molecule, e.g., Ab 67-11 (U.S. Provisional application No.61/645,453, U.S. Application Publication No. 2013/0302348, andInternational Application Publication No. WO 2013/169377), FI6 (U.S.Application Publication No. 2010/0080813), FI28 (U.S. ApplicationPublication No. 2010/0080813), C179 (Okuno et al., J. Virol. 67:2552-8,1993), F10 (Sui et al., Nat. Struct. Mol. Biol. 16:265, 2009), CR9114(Dreyfus et al., Science 337:1343, 2012), or CR6261 (Ekiert et al.,Science 324:246, 2009). Thus, the formulation described herein (e.g., aformulation comprising Ab 044) can be used in combination of any ofthose antibodies.

In an embodiment the second or additional agent comprises a second oradditional antibody molecule, e.g., an anti-HA antibody, e.g., ananti-HA antibody disclosed herein. For example, two or more of Ab 044,Ab 069, Ab 032, and Ab 031 can be administered. For example, Ab 044 canbe administered in combination with Ab 069 or Ab 032.

In the case of combinations, two agents can be administered as part ofthe same dosage unit or administered separately. Other exemplary agentsuseful for treating the symptoms associated with influenza infection areacetaminophen, ibuprofen, aspirin, and naproxen.

In an embodiment the formulation, e.g., pharmaceutical formulation, isadministered to a human subject suffering from or susceptible to aninfluenza infection. In an embodiment, the formulation, e.g.,pharmaceutical formulation, is administered prior to known exposure toinfluenza, or to particular influenza subtypes or strains. In anembodiment, the formulation, e.g., pharmaceutical formulation, isadministered prior to manifestation of effects or symptoms of influenzainfection, or to one or more particular effects manifestation of effectsor symptoms of influenza infection. In an embodiment, the formulation,e.g., pharmaceutical formulation, is administered after known exposureto influenza, or to particular influenza subtypes or strains. In anembodiment, the formulation, e.g., pharmaceutical formulation, isadministered after manifestation of effects or symptoms of influenzainfection, or after observation of one or more particular effectsmanifestation of effects or symptoms of influenza infection. In anembodiment, the formulation, e.g., pharmaceutical formulation, isadministered in response to, or to treat or prevent, a manifestation ofan effect or a symptom of influenza infection, e.g., inflammation,fever, nausea, weight loss, loss of appetite, rapid breathing, increaseheart rate, high blood pressure, body aches, muscle pain, eye pain,fatigue, malaise, dry cough, runny nose, and/or sore throat.

In an embodiment, the method further comprises, testing the subject,e.g., human subject, for the influenza virus, e.g., with a methoddisclosed herein. In an embodiment, the administration is responsive toa positive test for influenza.

In yet another aspect, the disclosure features a method of treating asubject, e.g., a human subject, infected with an influenza virus (e.g.,an influenza A virus, e.g., a Group 1 strain, e.g., an H1N1 strain,e.g., A/South Carolina/1/1918, A/Puerto Rico/08/1934, orA/California/04/2009, or an H5N1 strain, e.g., A/Indonesia/5/2005 orA/Vietnam/1203/2004, or an influenza B virus, e.g., B/Wisconsin/1/2010)by administering a formulation, e.g., pharmaceutical formulation,described herein. For example, the influenza A virus is an H1, H5, H9,H3 or H7 strain, such as an H1N1 strain, an H3N2 strain, an H5N1 strain,or an H7N9 strain of influenza A virus.

In an embodiment, the formulation, e.g., pharmaceutical formulation, isadministered instead of a vaccine for prevention of influenza. Inanother embodiment, the formulation, e.g., pharmaceutical formulation,is administered in combination with (simultaneously or sequentiallywith) a vaccine for prevention of the influenza.

In yet another aspect, the disclosure features a method of detectinginfluenza (e.g., influenza A or influenza B) virions in a biologicalsample, such as by contacting the sample with a formulation, e.g.,pharmaceutical formulation, comprising an anti-HA antibody moleculedescribed herein, and then detecting the binding of the antibodymolecule to the sample. In an embodiment, the method of detecting theinfluenza virus (e.g., influenza A or influenza B virus) is performed invitro.

In one aspect, the disclosure features a method of (a) providing asample from a patient; (b) contacting the sample with a formulation,e.g., pharmaceutical formulation, comprising an anti-HA antibodymolecule described herein, and (c) determining whether the antibodymolecule binds a polypeptide in the sample, where if the antibodymolecule binds a polypeptide in the sample, then the patient isdetermined to be infected with an influenza virus (e.g., an influenza Avirus, e.g., a Group 1 strain, e.g., an H1N1 strain, e.g., A/SouthCarolina/1/1918, A/Puerto Rico/08/1934, or A/California/04/2009, or anH5N1 strain, e.g., A/Indonesia/5/2005 or A/Vietnam/1203/2004, or aninfluenza B virus, e.g., B/Wisconsin/1/2010). In an embodiment, thepatient is determined to be infected with an influenza virus (e.g., aninfluenza A virus, e.g., a Group 1 strain, e.g., an H1N1 strain, e.g.,A/South Carolina/1/1918, A/Puerto Rico/08/1934, or A/California/04/2009,or an H5N1 strain, e.g., A/Indonesia/5/2005 or A/Vietnam/1203/2004, oran influenza B virus, e.g., B/Wisconsin/1/2010), and the patient isfurther administered a formulation or an antibody molecule, disclosedherein, with which the test was performed.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Although methods and materialssimilar or equivalent to those described herein can be used in thepractice or testing of the invention, suitable methods and materials aredescribed below. All publications, patent applications, patents, andother references mentioned herein are incorporated by reference in theirentirety. In case of conflict, the present specification, includingdefinitions, will control. In addition, the materials, methods, andexamples are illustrative only and not intended to be limiting.

The details of one or more embodiments featured in the disclosure areset forth in the accompanying drawings and the description below. Otherfeatures, objects, and advantages featured in the disclosure will beapparent from the description and drawings, and from the claims.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is the heavy and light chain amino acid sequences (SEQ ID NOs:94and 95, respectively) of the anti-HA antibody A18. The constant domainsequence is indicated by italics. The CDRs are indicated by underlining.

FIG. 2 is the variable heavy chain domain sequence of exemplary anti-HAantibodies. The SEQ ID NOs for sequences shown are as follows: VH15 isSEQ ID NO: 15; VH16 is SEQ ID NO: 16; VH17 is SEQ ID NO: 17; VH18 is SEQID NO: 18; VH19 is SEQ ID NO: 19; VH21 is SEQ ID NO: 21; VH22 is SEQ IDNO: 22; VH20 is SEQ ID NO: 20; VH23 is SEQ ID NO: 23; VH24 is SEQ ID NO:24; VH25 is SEQ ID NO: 25; VH26 is SEQ ID NO: 26; VH27 is SEQ ID NO: 27;and VH161 is SEQ ID NO: 161.

FIGS. 3A-3B depict the variable light chain domain sequence of exemplaryanti-HA antibodies. The SEQ ID NOs for sequences shown are as follows:VL28 is SEQ ID NO: 28; VL29 is SEQ ID NO: 29; VL30 is SEQ ID NO: 30;VL35 is SEQ ID NO: 35; VL31 is SEQ ID NO: 31; VL32 is SEQ ID NO: 32;VL33 is SEQ ID NO: 33; VL34-ID is SEQ ID NO: 34; VL36 is SEQ ID NO: 36;VL45 is SEQ ID NO: 45; VL46 is SEQ ID NO: 46; VL37 is SEQ ID NO: 37;VL38 is SEQ ID NO: 38; VL39 is SEQ ID NO: 39; VL40 is SEQ ID NO: 40;VL41 is SEQ ID NO: 41; VL42 is SEQ ID NO: 42; VL43 is SEQ ID NO: 43;VL44 is SEQ ID NO: 44; VL47 is SEQ ID NO: 47; VL48 is SEQ ID NO: 48;VL49 is SEQ ID NO: 49; VL50 is SEQ ID NO: 50; VLSI is SEQ ID NO: 51;VL52 is SEQ ID NO: 52; VL53 is SEQ ID NO: 53; VL54 is SEQ ID NO: 54;VL55 is SEQ ID NO: 55; VL56 is SEQ ID NO: 56; VL57 is SEQ ID NO: 57;VL58 is SEQ ID NO: 58; VL59 is SEQ ID NO: 59; VL60 is SEQ ID NO: 60;VL61 is SEQ ID NO: 61; VL153 is SEQ ID NO: 153; VL154 is SEQ ID NO: 154;VL155 is SEQ ID NO: 155; VL156 is SEQ ID NO: 156; and VL62 is SEQ ID NO:62.

FIG. 4 shows the amino acid sequences of the heavy chain variableregions of FI6 (SEQ ID NO: 175), FI370 (SEQ ID NO: 176), FI6 variant 1(SEQ ID NO: 177), FI6 variant 3 (SEQ ID NO: 178), FI6/370 (SEQ ID NO:179) and the amino acid sequence of kappa light chain variable region ofFI6 (SEQ ID NO: 180).

FIG. 5 is the variable heavy chain domain sequence of exemplary anti-HAantibodies as shown in FIG. 2 and including an N-terminal ID dipeptide.The SEQ ID NOs. for sequences shown are as follows: VH15-ID is SEQ IDNO: 96; VH16-ID is SEQ ID NO: 97; VH17-ID is SEQ ID NO: 98; VH18-ID isSEQ ID NO: 99; VH19-ID is SEQ ID NO: 100; VH21-ID is SEQ ID NO: 101;VH22-ID is SEQ ID NO: 102; VH20-ID is SEQ ID NO: 103; VH23-ID is SEQ IDNO: 104; VH24-ID is SEQ ID NO: 105; VH25-ID is SEQ ID NO: 106; VH26-IDis SEQ ID NO: 107; VH27-ID is SEQ ID NO: 108; and VH161-ID is SEQ ID NO:109.

FIGS. 6A-6B depict the variable light chain domain sequence of exemplaryanti-HA antibodies as shown in FIGS. 3A-3B and including an N-terminalID dipeptide. The SEQ ID NOs for sequences shown are as follows: VL28-IDis SEQ ID NO: 110; VL29-ID is SEQ ID NO: 111; VL30-ID is SEQ ID NO: 112;VL35-ID is SEQ ID NO: 113; VL31-ID is SEQ ID NO: 114; VL32-ID is SEQ IDNO: 115; VL33-ID is SEQ ID NO: 116; VL34-ID is SEQ ID NO: 117; VL36-IDis SEQ ID NO: 118; VL45-ID is SEQ ID NO: 119; VL46-ID is SEQ ID NO: 120;VL37-ID is SEQ ID NO: 121; VL38-ID is SEQ ID NO: 122; VL39-ID is SEQ IDNO: 123; VL40-ID is SEQ ID NO: 124; VL41-ID is SEQ ID NO: 125; VL42-IDis SEQ ID NO: 126; VL43-ID is SEQ ID NO: 127; VL44-ID is SEQ ID NO: 128;VL47-ID is SEQ ID NO: 129; VL48-ID is SEQ ID NO: 130; VL49-ID is SEQ IDNO: 131; VL50-ID is SEQ ID NO: 132; VL51-ID is SEQ ID NO: 133; VL52-IDis SEQ ID NO: 134; VL53-ID is SEQ ID NO: 135; VL54-ID is SEQ ID NO: 136;VL55-ID is SEQ ID NO: 137; VL56-ID is SEQ ID NO: 138; VL57-ID is SEQ IDNO: 139; VL58-ID is SEQ ID NO: 140; VL59-ID is SEQ ID NO: 141; VL60-IDis SEQ ID NO: 142; VL61-ID is SEQ ID NO: 143; VL153-ID is SEQ ID NO:157; VL154-ID is SEQ ID NO: 158; VL155-ID is SEQ ID NO: 159; VL156-ID isSEQ ID NO: 160; and VL62-ID is SEQ ID NO: 144.

FIG. 7 shows the variable light and heavy chain sequences of additionalexemplary anti-HA antibodies. The SEQ ID NOs for sequences shown are asfollows: VL165 is SEQ ID NO: 165; VL166 is SEQ ID NO: 166; VL167 is SEQID NO: 167; VL168 is SEQ ID NO: 168; VL169 is SEQ ID NO: 169; VH164 isSEQ ID NO: 164; VH162 is SEQ ID NO: 162; VH163 is SEQ ID NO: 163.

FIGS. 8A-8G show the DSC profile for all of the 14 formulation samplestested in Example 2.

FIG. 9 depicts representative reduced CE-SDS electropherogram(Formulation #1, 45° C., 2 wks).

FIG. 10 depicts representative non-reduced CE-SDS electropherogram(Formulation #1, 45° C., 2 wks).

FIG. 11 depicts representative SEC chromatogram (Formulation #1, 45° C.,2 wks).

FIG. 12 depicts a representative IEF gel image.

DETAILED DESCRIPTION

The disclosure is based, at least in part, on the design and synthesisof antibody molecules that can bind an epitope that is conserved acrossmultiple hemagglutinin subtypes of influenza viruses (e.g., influenza Aand influenza B viruses). For example, formulations (e.g.,pharmaceutical formulations) comprising the antibody molecules describedherein are useful as broad spectrum therapy against disease caused by atleast one influenza A strain belonging to Group 1 and one influenza Astrain belonging to Group 2 to neutralize infectivity of virusesbelonging to both Group 1 and Group 2 (at least one subtype of each).Without wishing to be bound by theory, it is belived that theformulations (e.g., pharmaceutical formulations) described herein aresuitable for use in treating or preventing influenza viruses, at leastin part, because the formulations (e.g., pharmaceutical formulations)have one or more desired properties such as improved stability (e.g.,low degradation and/or aggregation) and maintained potency (e.g., HAbinding).

The antibody molecules were designed by a rational structure-basedapproach to target a region on the virus that is not fully accessible tothe human immune system and, therefore, not amenable to antibodyselection through more classical screening approaches. Thisrational-based approach to the design and development of broad-spectrumantibody molecules allows for the development of more efficaciousvaccines for pandemic and seasonal influenza. This approach also allowsfor the advance preparation of pandemic vaccines so that they are readyto be employed against specific virus subtypes (e.g., avian virussubtypes) that may mutate to become human-adapted and highlytransmissible. Vaccines (e.g., seasonal vaccines) that utilize theapproach described herein can generate a more potent immune responsewithout the use of adjuvants and provide broad protection against viralstrain variation.

Definitions

As used herein, the term “antibody molecule” refers to a polypeptidethat comprises sufficient sequence from an immunoglobulin heavy chainvariable region and/or sufficient sequence from an immunoglobulin lightchain variable region, to provide antigen specific binding. It comprisesfull length antibodies as well as fragments thereof, e.g., Fabfragments, that support antigen binding. Typically an antibody moleculewill comprise heavy chain CDR1, CDR2, and CDR3 and light chain CDR1,CDR2, and CDR3 sequence. Antibody molecules include human, humanized,CDR-grafted antibodies and antigen binding fragments thereof. In anembodiment, an antibody molecule comprises a protein that comprises atleast one immunoglobulin variable region segment, e.g., an amino acidsequence that provides an immunoglobulin variable domain orimmunoglobulin variable domain sequence.

The VH or VL chain of the antibody molecule can further include all orpart of a heavy or light chain constant region, to thereby form a heavyor light immunoglobulin chain, respectively. In an embodiment, theantibody molecule is a tetramer of two heavy immunoglobulin chains andtwo light immunoglobulin chains.

An antibody molecule can comprise one or both of a heavy (or light)chain immunoglobulin variable region segment. As used herein, the term“heavy (or light) chain immunoglobulin variable region segment,” refersto an entire heavy (or light) chain immunoglobulin variable region, or afragment thereof, that is capable of binding antigen. The ability of aheavy or light chain segment to bind antigen is measured with thesegment paired with a light or heavy chain, respectively. In someembodiment, a heavy or light chain segment that is less than a fulllength variable region will, when paired with the appropriate chain,bind with an affinity that is at least 20, 30, 40, 50, 60, 70, 80, 90,or 95% of what is seen when the full length chain is paired with a lightchain or heavy chain, respectively.

An immunoglobulin variable region segment may differ from a reference orconsensus sequence. As used herein, to “differ,” means that a residue inthe reference sequence or consensus sequence is replaced with either adifferent residue or an absent or inserted residue.

An antibody molecule can comprise a heavy (H) chain variable region(abbreviated herein as VH), and a light (L) chain variable region(abbreviated herein as VL). In another example, an antibody comprisestwo heavy (H) chain variable regions and two light (L) chain variableregions or antibody binding fragments thereof. The light chains of theimmunoglobulin may be of types kappa or lambda. In an embodiment, theantibody molecule is glycosylated. An antibody molecule can befunctional for antibody dependent cytotoxicity and/orcomplement-mediated cytotoxicity, or may be non-functional for one orboth of these activities. An antibody molecule can be an intact antibodyor an antigen-binding fragment thereof.

Antibody molecules include “antigen-binding fragments” of a full lengthantibody, e.g., one or more fragments of a full-length antibody thatretain the ability to specifically bind to an HA target of interest.Examples of binding fragments encompassed within the term“antigen-binding fragment” of a full length antibody include (i) a Fabfragment, a monovalent fragment consisting of the VL, VH, CL and CH1domains; (ii) a F(ab′) or F(ab′)₂ fragment, a bivalent fragmentincluding two Fab fragments linked by a disulfide bridge at the hingeregion; (iii) an Fd fragment consisting of the VH and CH1 domains; (iv)an Fv fragment consisting of the VL and VH domains of a single arm of anantibody, (v) a dAb fragment (Ward et al., (1989) Nature 341:544-546),which consists of a VH domain; and (vi) an isolated complementaritydetermining region (CDR) that retains functionality. Furthermore,although the two domains of the Fv fragment, VL and VH, are coded for byseparate genes, they can be joined, using recombinant methods, by asynthetic linker that enables them to be made as a single protein chainin which the VL and VH regions pair to form monovalent molecules knownas single chain Fv (scFv). See e.g., Bird et al. (1988) Science242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA85:5879-5883. Antibody molecules include diabodies.

As used herein, an antibody refers to a polypeptide, e.g., a tetramericor single chain polypeptide, comprising the structural and functionalcharacteristics, particularly the antigen binding characteristics, of animmunoglobulin. Typically, a human antibody comprises two identicallight chains and two identical heavy chains. Each chain comprises avariable region.

The variable heavy (VH) and variable light (VL) regions can be furthersubdivided into regions of hypervariability, termed “complementaritydetermining regions” (“CDR”), interspersed with regions that are moreconserved, termed “framework regions” (FR). Human antibodies have threeVH CDRs and three VL CDRs, separated by framework regions FR1-FR4. Theextent of the FRs and CDRs has been precisely defined (see, Kabat, E.A., et al. (1991) Sequences of Proteins of Immunological Interest, FifthEdition, U.S. Department of Health and Human Services, NIH PublicationNo. 91-3242; and Chothia, C. et al. (1987) J. Mol. Biol. 196:901-917).Kabat definitions are used herein. Each VH and VL is typically composedof three CDRs and four FRs, arranged from amino-terminus tocarboxyl-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3,CDR3, FR4.

The heavy and light immunoglobulin chains can be connected by disulfidebonds. The heavy chain constant region typically comprises threeconstant domains, CHL CH2 and CH3. The light chain constant regiontypically comprises a CL domain. The variable region of the heavy andlight chains contains a binding domain that interacts with an antigen.The constant regions of the antibodies typically mediate the binding ofthe antibody to host tissues or factors, including various cells of theimmune system (e.g., effector cells) and the first component (Clq) ofthe classical complement system.

The term “immunoglobulin” comprises various broad classes ofpolypeptides that can be distinguished biochemically. Those skilled inthe art will appreciate that heavy chains are classified as gamma, mu,alpha, delta, or epsilon (γ, μ, α, δ, ε) with some subclasses among them(e.g., γ1-γ4). It is the nature of this chain that determines the“class” of the antibody as IgG, IgM, IgA IgD, or IgE, respectively. Theimmunoglobulin subclasses (isotypes) e.g., IgG1, IgG2, IgG3, IgG4, IgA1,etc. are well characterized and are known to confer functionalspecialization. Modified versions of each of these classes and isotypesare readily discernable to the skilled artisan in view of the instantdisclosure and, accordingly, are within the scope of the instantdisclosure. All immunoglobulin classes are clearly within the scope ofthe present disclosure. Light chains are classified as either kappa orlambda (κ, λ). Each heavy chain class may be bound with either a kappaor lambda light chain.

Suitable antibodies include, but are not limited to, monoclonal,monospecific, polyclonal, polyspecific, human antibodies, primatizedantibodies, chimeric antibodies, bi-specific antibodies, humanizedantibodies, conjugated antibodies (e.g., antibodies conjugated or fusedto other proteins, radiolabels, or cytotoxins), Small ModularImmunoPharmaceuticals (“SMIPs™”), single chain antibodies, cameloidantibodies, and antibody fragments.

In an embodiment, an antibody is a humanized antibody. A humanizedantibody refers to an immunoglobulin comprising a human framework regionand one or more CDR's from a non-human, e.g., mouse or rat,immunoglobulin. The immunoglobulin providing the CDR's is often referredto as the “donor” and the human immunoglobulin providing the frameworkoften called the “acceptor,” though in an embodiment, no source or noprocess limitation is implied. Typically a humanized antibody comprisesa humanized light chain and a humanized heavy chain immunoglobulin.

An “immunoglobulin domain” refers to a domain from the variable orconstant domain of immunoglobulin molecules. Immunoglobulin domainstypically contain two β-sheets formed of about seven β-strands, and aconserved disulfide bond (see e.g., A. F. Williams and A. N. Barclay(1988) Ann. Rev. Immunol. 6:381-405).

As used herein, an “immunoglobulin variable domain sequence” refers toan amino acid sequence that can form the structure of an immunoglobulinvariable domain. For example, the sequence may include all or part ofthe amino acid sequence of a naturally-occurring variable domain. Forexample, the sequence may omit one, two or more N- or C-terminal aminoacids, internal amino acids, may include one or more insertions oradditional terminal amino acids, or may include other alterations. In anembodiment, a polypeptide that comprises an immunoglobulin variabledomain sequence can associate with another immunoglobulin variabledomain sequence to form a target binding structure (or “antigen bindingsite”), e.g., a structure that interacts with the target antigen.

As used herein, the term antibodies comprises intact monoclonalantibodies, polyclonal antibodies, single domain antibodies (e.g., sharksingle domain antibodies (e.g., IgNAR or fragments thereof)),multispecific antibodies (e.g., bi-specific antibodies) formed from atleast two intact antibodies, and antibody fragments so long as theyexhibit the desired biological activity. Antibodies for use herein maybe of any type (e.g., IgA, IgD, IgE, IgG, or IgM).

The antibody or antibody molecule can be derived from a mammal, e.g., arodent, e.g., a mouse or rat, horse, pig, or goat. In an embodiment, anantibody or antibody molecule is produced using a recombinant cell. Insome embodiments an antibody or antibody molecule is a chimericantibody, for example, from mouse, rat, horse, pig, or other species,bearing human constant and/or variable regions domains.

A binding agent, as used herein, is an agent that bind, e.g.,specifically binds, a target antigen, e.g., HA. Binding agents of theinvention share sufficient structural relationship with anti-HA antibodymolecules disclosed herein to support specific binding to HA, and in anembodiment, other functional properties of an anti-HA antibody moleculedisclosed herein. In an embodiment, a binding agent will exhibit abinding affinity at of at least 10, 20, 30, 40, 50, 60, 70, 80, or 90%of an antibody molecule disclosed herein, e.g., an antibody moleculewith which it shares, significant structural homology, e.g., CDRsequences. Binding agents can be naturally occurring, e.g., as are someantibodies, or synthetic. In an embodiment a binding agents is apolypeptide, e.g., an antibody molecule, e.g., an antibody. While somebinding agents are antibody molecules, other molecules, e.g., otherpolypeptides, can also function as binding agents. Polypeptide bindingagents can be monomeric or multimeric, e.g., dimeric, trimeric, ortetrameric and can be stabilized by intra- or interchain bonds, e.g.,disulfide bonds. They can contain natural or non-naturally occurringamino acid residues. In an embodiment, binding agents are antibodymolecules, or other polypeptides, that present one or more CDRs ofantibody molecules disclosed herein or that otherwise mimic thestructure of an antibody molecule disclosed herein. Binding agents canalso comprise aptomers, nucleic acids or other molecular entities. Abinding agent can be developed in a variety of ways, e.g., byimmunization, by rational design, screening of random structures, or acombination of those or other approaches. Typically a binding agent willact by making contact with substantially the same epitope as an antibodymolecule disclosed herein, e.g., an antibody molecule with which itshares, significant structural homology, e.g., CDR sequences. A bindingagent can interact with amino acids, saccharides, or combinationsthereof. Polypeptides other than antibodies can be used as a scaffold topresent sequence, e.g., one or more, or a complete set of heavy chainand/or light chain CDRs, disclosed herein. Exemplary scaffolds includeadnectin, zinc finger DNA-binding proteins. protein A, lipoclins,ankryin consensus repeat domain, thioredoxin, anticalins, centyrin,avimer domains, ubiquitin, peptidomimetics, stapled peptides,cystine-knot miniproteins, and IgNARs. In some embodiments, a bindingagent is or comprises a nucleic acid, e.g., DNA, RNA or mixturesthereof. In an embodiment, a binding agent, e.g., a nucleic acid, showssecondary, tertiary, or quaternary structure. In some embodiments abinding agent, e.g., a nucleic acid, forms a structure that mimics thestructure of an antibody molecule disclosed herein.

A broad spectrum binding agent, e.g., antibody molecule, as used herein,binds, a plurality of different HA molecules, and optionally neutralizesviruses comprising the different HA molecules. In an embodiment it bindsa first HA and binds a second HA from influenza A Group 1, andoptionally neutralizes viruses comprising the first or second HAmolecules. In an embodiment, it binds a first HA from an influenza AGroup 1 virus, and binds a second HA from an influenza A Group 2 virus,and optionally neutralizes viruses comprising the different HAmolecules. In an embodiment it binds a first HA from an influenza AGroup 1 or 2 virus and binds a HA from an influenza B virus, andoptionally neutralizes viruses comprising the different HA molecules. Inan embodiment, it binds, and in an embodiment neutralizes, at least twodifferent clades or clusters of virus, e.g., from different Groups. Inan embodiment, it binds, and in an embodiment neutralizes, all orsubstantially all strains of Group 1 and/or Group 2 disclosed herein. Inan embodiment, a binding agent, e.g., antibody molecule, binds, and inan embodiment, neutralizes: at least one strain from the Group 1 H1,e.g., H1a or H1b, cluster and at least one strain from the Group 2 H3 orH7 cluster. In an embodiment, a binding agent, e.g., antibody molecule,binds, and in an embodiment, neutralizes: at least one strain from theGroup 1 H1, e.g., H1a or H1b, cluster and at least one influenza Bstrain. In an embodiment, a binding agent, e.g., antibody molecule,binds, and in an embodiment, neutralizes: at least one strain from theGroup 2 H3 or H7 cluster and at least one influenza B strain. In anembodiment, a binding agent, e.g., antibody molecule, binds, and in anembodiment, neutralizes: at least one strain from the Group 1 H1, e.g.,H1a or H1b, cluster, at least one strain from the Group 2 H3 or H7cluster, and at least one influenza B strain. In some embodiments,binding agent, e.g., antibody molecule, binds, and optionallyneutralizes or mediate infection of particular hosts, e.g., avian,camel, canine, cat, civet, equine, human, mouse, swine, tiger, or othermammal or bird.

The term “combination therapy”, as used herein, refers to administrationof a plurality of agents, e.g., wherein at least one binding agent,e.g., antibody molecule, disclosed herein is administered to a subject,e.g., a human subject. The introduction of the agents into the subjectcan be at different times. In an embodiment, the agents are administeredin overlapping regimens, or such that the subject is simultaneouslyexposed to both agents, or such that the response of the subject isbetter than would be seen with either agent administered alone.

As used herein, an “escape mutant” is a mutated influenza strain that isresistant to neutralization by an anti-HA antibody molecule describedherein. In an embodiment, an escape mutant is resistant toneutralization with a binding agent, e.g., antibody molecule, but itsparent strain is neutralized by the binding agent, e.g., antibodymolecule.

As used herein, “pandemic influenza” refers to a new viral strain thatarises due to human adaptation of an influenza strain by mutation or byemergence of a strain by reassortment of different strains of influenzaA. The resulting pandemic strain is significantly different fromprevious strains and most people will have little or no pre-existingimmunity Symptoms and complications may be more severe and more frequentthan those typical of seasonal influenza. Examples of past pandemic fluviruses include, e.g., the 2009 H1N1 ‘swine flu,’ the 1957-58 H2N2‘Asian flu’ and the 1968 H3N2 influenza strains.

The terms “purified” and “isolated” as used herein in the context of anantibody molecule, e.g., a antibody, a immunogen, or generally apolypeptide, obtained from a natural source, refers to a molecule whichis substantially free of contaminating materials from the naturalsource, e.g., cellular materials from the natural source, e.g., celldebris, membranes, organelles, the bulk of the nucleic acids, orproteins, present in cells. Thus, a polypeptide, e.g., an antibodymolecule, that is isolated includes preparations of a polypeptide havingless than about 30%, 20%, 10%, 5%, 2%, or 1% (by dry weight) of cellularmaterials and/or contaminating materials. The terms “purified” and“isolated” when used in the context of a chemically synthesized species,e.g., an antibody molecule, or immunogen, refers to the species which issubstantially free of chemical precursors or other chemicals which areinvolved in the syntheses of the molecule.

A preparation of binding agents, e.g., antibody molecules, as usedherein, comprises a plurality of molecules of a binding agent, e.g.,antibody molecule, described herein. In an embodiment, that bindingagent, e.g., antibody molecule, makes up at least 60, 70, 80, 90, 95,98, 99, 99.5 or 99.9%, of the preparation, or of the active ingredientsof the preparation, by weight or number. In an embodiment, that bindingagent is an antibody molecule which makes up at least 60, 70, 80, 90,95, 98, 99, 99.5 or 99.9%, of the preparation, or of the activeingredients, or polypeptide ingredients, or antibody molecules, of thepreparation, by weight or number. In an embodiment, the binding agent isan antibody molecule and the preparation contains no more than 30, 20,10, 5, 2, 1, or 0.5%, by weight or number, of a contaminant, e.g., areactant, solvent, precursor or other species, from the source, or usedin the preparation, of the antibody molecule, e.g., a species from acell, reaction mixture, or other system used to produce the antibodymolecule.

As used herein, the term “prevent infection” means that a subject (e.g.,a human) is less likely to be infected by influenza if the subjectreceives the antibody prior to (e.g., 1 day, 2 days, 1 week, 2 weeks, 3weeks, or 1 month of more) before being exposed to influenza.

As used herein, “seasonal influenza” is a strain that is identical orclosely related to strains that have been circulating in the humanpopulation in recent years and therefore most people are at leastpartially immune to it. Such a strain is not likely to cause severedisease. Symptoms can include fever, cough, runny nose, and muscle pain,and in rare cases, death can result from complications, such aspneumonia. Outbreaks follow predictable seasonal patterns, annually, andusually in fall and winter and in temperate climates. Infection due toseasonal influenza is commonly referred to as the flu.

As used herein, specific binding, means that a binding agent, e.g., anantibody molecule, binds its antigen with a K_(D) of equal to or lessthan 10⁻⁵ nM. In an embodiment, the antibody binds it's antigen with aK_(D) of equal to or less than 10⁻⁶, 10⁻⁷, 10⁻⁸, 10⁻⁹, 10⁻¹⁰, 10⁻¹¹, or10⁻¹² nM.

As used herein, the term “therapeutically effective amount” refers to anamount of a therapeutic agent, e.g., a binding agent, e.g., an antibodymolecule, which results in a positive outcome for the subject. In anembodiment, it can be statistically correlated with therapeutic effector benefit, e.g., the lessening or prevention of a manifestation of aneffect or a symptom, when administered to a population of subjects. Inan embodiment, it is an amount that also provides a preselected, orreasonable, benefit/risk ratio. In an embodiment, it is an amounteffective to reduce the incidence and/or severity of and/or to delayonset of one or more features, symptoms, or characteristics of adisease, disorder, or condition. A therapeutically effective amount iscan be administered in a dosing regimen that may comprise one ormultiple unit doses.

As used herein, the term “treat infection” means that a subject (e.g., ahuman) who has been infected with an influenza and experiences symptomsof the influenza (e.g., the flu), will In an embodiment, suffer lesssevere symptoms and/or will recover faster when the antibody molecule isadministered than if the antibody is never administered. In anembodiment, when an infection is treated, an assay to detect virus inthe subject will detect less virus after effective treatment for theinfection. For example, a diagnostic assay using an antibody molecule,such as an antibody molecule described herein, will detect less or novirus in a biological sample of a patient after administration of anantibody molecule for the effective treatment of the viral infection.Other assays, such as PCR (e.g., qPCR) can also be used to monitortreatment in a patient, to detect the presence, e.g., decreased presence(or absence) after treatment of viral infection in the patient.Treatment can, e.g., partially or completely alleviate, ameliorate,relive, inhibit, reduce the severity of, and/or reduces incidence andoptionally, delay onset of, one or more manifestations of the effects orsymptoms, features, and/or causes of a particular disease, disorder,and/or condition (e.g., influenza). In an embodiment, treatment is of asubject who does not exhibit signs of the relevant disease, disorderand/or condition and/or of a subject who exhibits only early signs ofthe disease, disorder, and/or condition. In an embodiment, treatment isof a subject who exhibits one or more established signs of the relevantdisease, disorder and/or condition. In an embodiment, treatment is of asubject diagnosed as suffering from influenza.

Calculations of “homology” or “sequence identity” or “identity” betweentwo sequences (the terms are used interchangeably herein) can beperformed as follows. The sequences are aligned for optimal comparisonpurposes (e.g., gaps can be introduced in one or both of a first and asecond amino acid or nucleic acid sequence for optimal alignment andnon-homologous sequences can be disregarded for comparison purposes).The optimal alignment is determined as the best score using the GAPprogram in the GCG software package with a Blossum 62 scoring matrixwith a gap penalty of 12, a gap extend penalty of 4, and a frameshiftgap penalty of 5. The amino acid residues or nucleotides atcorresponding amino acid positions or nucleotide positions are thencompared. When a position in the first sequence is occupied by the sameamino acid residue or nucleotide as the corresponding position in thesecond sequence, then the molecules are identical at that position (asused herein amino acid or nucleic acid “identity” is equivalent to aminoacid or nucleic acid “homology”). The percent identity between the twosequences is a function of the number of identical positions shared bythe sequences.

Formulations

The binding agents, e.g., antibody molecules, described herein can beformulated, e.g., as pharmaceutical compositions, such as for thetreatment or prevention of influenza.

Typically, a pharmaceutical composition includes a pharmaceuticallyacceptable carrier. As used herein, “pharmaceutically acceptablecarrier” includes any and all solvents, dispersion media, coatings,antibacterial and antifungal agents, isotonic and absorption delayingagents, and the like that are physiologically compatible.

A “pharmaceutically acceptable salt” refers to a salt that retains thedesired biological activity of the parent compound and does not impartany undesired toxicological effects (see e.g., Berge, S. M., et al.(1977) J. Pharm. Sci. 66:1-19). Examples of such salts include acidaddition salts and base addition salts. Acid addition salts includethose derived from nontoxic inorganic acids, such as hydrochloric,nitric, phosphoric, sulfuric, hydrobromic, hydroiodic, and the like, aswell as from nontoxic organic acids such as aliphatic mono- anddicarboxylic acids, phenyl-substituted alkanoic acids, hydroxy alkanoicacids, aromatic acids, aliphatic and aromatic sulfonic acids and thelike. Base addition salts include those derived from alkaline earthmetals, such as sodium, potassium, magnesium, calcium and the like, aswell as from nontoxic organic amines, such asN,N′-dibenzylethylenediamine, N-methylglucamine, chloroprocaine,choline, diethanolamine, ethylenediamine, procaine and the like.

The compositions comprising the antibody molecules described herein canbe formulated according to methods known in the art. Pharmaceuticalformulation is a well-established art, and is further described inGennaro (ed.), Remington: The Science and Practice of Pharmacy, 20^(th)ed., Lippincott, Williams & Wilkins (2000) (ISBN: 0683306472); Ansel etal., Pharmaceutical Dosage Forms and Drug Delivery Systems, 7^(th) Ed.,Lippincott Williams & Wilkins Publishers (1999) (ISBN: 0683305727); andKibbe (ed.), Handbook of Pharmaceutical Excipients AmericanPharmaceutical Association, 3^(rd) ed. (2000) (ISBN: 091733096X).

Pharmaceutical compositions may be in a variety of forms. These include,for example, liquid, semi-solid and solid dosage forms, such as liquidsolutions (e.g., injectable and infusible solutions), dispersions orsuspensions, tablets, pills, powders, liposomes and suppositories. Theform can depend on the intended mode of administration and therapeuticapplication. Typically, compositions for the agents described herein arein the form of injectable or infusible solutions.

Such compositions can be administered by a parenteral mode (e.g.,intravenous, subcutaneous, intraperitoneal, or intramuscular injection).The phrases “parenteral administration” and “administered parenterally”as used herein mean modes of administration other than enteral andtopical administration, usually by injection, and include, withoutlimitation, intravenous, intramuscular (IM), intraarterial, intrathecal,intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal,transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular,subarachnoid, intraspinal, epidural and by intrasternal injection or byinfusion.

Pharmaceutical compositions may be provided in a sterile injectable form(e.g., a form that is suitable for subcutaneous injection or intravenousinfusion). In an embodiment, the pharmaceutical composition is providedin a liquid dosage form that is suitable for injection or topicalapplication. In some embodiments, pharmaceutical compositions areprovided as in dry form, e.g., as powders (e.g. lyophilized and/orsterilized preparations). The Pharmaceutical composition can be providedunder conditions that enhance stability, e.g., under nitrogen or undervacuum. Dry material can be reconstituted with an aqueous diluent (e.g.,water, buffer, salt solution, etc.) prior to injection.

In an embodiment, the pharmaceutical composition containing an anti-HAantibody is administered intranasally. In another embodiment, thepharmaceutical composition containing an anti-HA antibody isadministered by inhalation, such as by oral or by nasal inhalation.

In an embodiment, the pharmaceutical composition is suitable for buccal,oral or nasal delivery, e.g., as a liquid, spray, or aerosol, e.g., bytopical application, e.g., by a liquid or drops, or by inhalation). Inan embodiment, a pharmaceutical preparation comprises a plurality ofparticles, suitable, e.g., for inhaled or aerosol delivery. In anembodiment, the mean particle size of 4, 5, 6, 7, 8, 9, 10, 11, 12, or13 microns. In an embodiment, a pharmaceutical preparation is formulatedas a dry powder, suitable, e.g., for inhaled or aerosol delivery. In anembodiment, a pharmaceutical preparation is formulated as a wet powder,through inclusion of a wetting agent, e.g., water, saline, or otherliquid of physiological pH. In an embodiment, a pharmaceuticalpreparation is provided as drops, suitable, e.g., for delivery to thenasal or buccal cavity.

In an embodiment, the pharmaceutical composition is disposed in adelivery device, e.g., a syringe, a dropper or dropper bottle, aninhaler, or a metered dose device, e.g., an inhaler. In an embodiment,the pharmaceutical composition is disposed in a container, e.g., anintravenous (IV) solution bag.

In an embodiment, a pharmaceutical composition contains a vector, suchas an adenovirus-associated virus (AAV)-based vector, that encodes aheavy chain of an anti-HA antibody molecule, and a light chain of ananti-HA antibody molecule, described herein. The composition containingthe vector can be administered to a subject, such as a patient, such asby injection, e.g., IM injection. Genes encoding the anti-HA antibodyunder control of, for example, cytomegalovirus (CMV) promoters, areexpressed in the body, and the recombinant anti-HA antibody molecule isintroduced into the circulation. See e.g., Balazs et al., Nature30:481:81-84, 2011.

Pharmaceutical compositions typically should be sterile and stable underthe conditions of manufacture and storage. A pharmaceutical compositioncan also be tested to insure it meets regulatory and industry standardsfor administration.

The composition can be formulated as a solution, microemulsion,dispersion, liposome, or other ordered structure suitable to high drugconcentration. Sterile injectable solutions can be prepared byincorporating an agent described herein in the required amount in anappropriate solvent with one or a combination of ingredients enumeratedabove, as required, followed by filtered sterilization. Generally,dispersions are prepared by incorporating an agent described herein intoa sterile vehicle that contains a basic dispersion medium and therequired other ingredients from those enumerated above. In the case ofsterile powders for the preparation of sterile injectable solutions,typical methods of preparation are vacuum drying and freeze-drying thatyields a powder of an agent described herein plus any additional desiredingredient from a previously sterile-filtered solution thereof. Theproper fluidity of a solution can be maintained, for example, by the useof a coating such as lecithin, by the maintenance of the requiredparticle size in the case of dispersion and by the use of surfactants.Prolonged absorption of injectable compositions can be brought about byincluding in the composition an agent that delays absorption, forexample, monostearate salts and gelatin.

A pharmaceutical composition may be provided, prepared, packaged, and/orsold in bulk, as a single unit dose, and/or as a plurality of singleunit doses. Typically a bulk preparation will contain at least 2, 5, 10,20, 50, or 100 unit doses. A unit dose is typically the amountintroduced into the patient in a single administration. In anembodiment, only a portion of a unit dose is introduced. In anembodiment, a small multiple, e.g., as much as 1.5, 2, 3, 5, or 10 timesa unit dose is administered. The amount of the active ingredient isgenerally equal to a dose which would be administered to a subjectand/or a convenient fraction of such a dose such as, for example,one-half or one-third of such a dose.

A formulation of a binding agent, e.g., an antibody molecule, caninclude, e.g., an anti-HA antibody molecule described herein, a buffer,and a tonicity agent. The pH of the formulation is generally pH 5.5-7.0.

In some embodiments, the formulation is a liquid formulation. In someembodiments, the formulation is stored as a liquid. In otherembodiments, the formulation is a lyophilized formulation. In certainembodiments, the formulation is prepared as a liquid and then is dried,e.g., by lyophilization or spray-drying, prior to storage. A driedformulation can be used as a dry compound, e.g., as an aerosol orpowder, or reconstituted to its original or another concentration, e.g.,using water, a buffer, or other appropriate liquid.

A “reconstituted” formulation is one which has been prepared bydissolving a lyophilized protein formulation in a diluent such that theprotein is dispersed in the reconstituted formulation. The reconstitutedformulation in suitable for administration (e.g. parenteraladministration) to a patient to be treated with the protein of interestand, in certain embodiments of the invention, may be one which issuitable for subcutaneous administration. The “diluent” of interestherein is one which is pharmaceutically acceptable (safe and non-toxicfor administration to a human) and is useful for the preparation of areconstituted formulation. Exemplary diluents include sterile water,bacteriostatic water for injection (BWFI), a pH buffered solution (e.g.phosphate-buffered saline), sterile saline solution, Ringer's solutionor dextrose solution.

A “lyoprotectant” is a molecule which, when combined with a protein ofinterest, significantly prevents or reduces chemical and/or physicalinstability of the protein upon lyophilization and subsequent storage.Exemplary lyoprotectants include sugars such as sucrose or trehalose; anamino acid such as monosodium glutamate or histidine; a methylamine suchas betaine; a lyotropic salt such as magnesium sulfate; a polyol such astrihydric or higher sugar alcohols, e.g. glycerin, erythritol, glycerol,arabitol, xylitol, sorbitol, and mannitol; propylene glycol;polyethylene glycol; pluronics; and combinations thereof. Typically, thelyoprotectant is a non-reducing sugar, such as trehalose or sucrose. Thelyoprotectant is added to the pre-lyophilized formulation in a“lyoprotecting amount” which means that, following lyophilization of theprotein in the presence of the lyoprotecting amount of thelyoprotectant, the protein essentially retains its physical and chemicalstability and integrity upon lyophilization and storage.

A “bulking agent” is a compound which adds mass to the lyophilizedmixture and contributes to the physical structure of the lyophilizedcake (e.g. facilitates the production of an essentially uniformlyophilized cake which maintains an open pore structure). Exemplarybulking agents include mannitol, glycine, polyethylene glycol andxorbitol.

In some embodiments, the anti-HA antibody molecule purification processis designed to permit transfer of an anti-HA antibody molecule into aformulation suitable for storage as a liquid. In other embodiments, theanti-HA antibody molecule purification process is designed to permittransfer of an anti-HA antibody molecule into a formulation forlong-term storage as a frozen liquid and subsequently for freeze-drying.

In some embodiments, the formulation is lyophilized with the protein ata specific concentration. The lyophilized formulation can then bereconstituted as needed with a suitable diluent (e.g., water) toresolubilize the original formulation components to a desiredconcentration, generally the same or higher concentration compared tothe concentration prior to lyophilization. The lyophilized formulationmay be reconstituted to produce a formulation that has a concentrationthat differs from the original concentration (e.g., beforelyophilization), depending upon the amount of water or diluent added tothe lyophilate relative to the volume of liquid that was originallyfreeze-dried. Suitable formulations can be identified by assaying one ormore parameters of antibody integrity. The assayed parameters aregenerally the percentage of HMW species or the percentage of LMWspecies.

The percentage of HMW species or LMW species is determined either as apercentage of the total protein content in a formulation or as a changein the percentage increase over time (e.g., during storage). The totalpercentage of HMW species in an acceptable formulation is not greaterthan 10% (e.g., not greater than 5%, not greater than 4%, not greaterthan 3%, not greater than 2%, or not greater than 1%) HMW species afterstorage as a lyophilate or liquid at 2° C. to 50° C. (e.g., at 4° C. to45° C., at 4° C. to 25° C., at 4° C. to 15° C., at about 4° C., at about25° C., or at about 45° C.) for at least one week, two weeks, one month,three months, six months, nine months, or one year or not greater thanabout 10% LMW species after storage as a lyophilate or liquid at 2° C.to 50° C. (e.g., at 4° C. to 45° C., at 4° C. to 25° C., at 4° C. to 15°C., at about 4° C., at about 25° C., or at about 45° C.) for at leastone week, two weeks, one month, three months, six months, nine months,or one year. In an embodiment, the total percentage of HMW species isnot greater than 5%. In another embodiment, the total percentage of HMWspecies is not greater than 3%. By “about” is meant ±20% of a citednumerical value. Thus, for example, “about 20° C.” means 16° C. to 24°C.

Typically, the stability profile is less than 10% HMW/LMW at 2°−8° C.for a refrigerated product, and 25° C. for a room-temperature product.HMW species or LMW species are assayed in a formulation stored as alyophilate after the lyophilate is reconstituted. 45° C. is anaccelerated condition that is generally used for testing stability anddetermining stability for short-term exposures to non-storageconditions, e.g., as may occur during transfer of a product duringshipping.

When the assayed parameter is the percentage change in HMW species orLMW species, the percent of total protein in one or both species afterstorage is compared to the percent total protein in one or both speciesprior to storage (e.g., upon preparation of the formulation). Thedifference in the percentages is determined. In general, the change inthe percentage of protein in HMW species or LMW species in liquidformulations is not greater than 10%, e.g., not greater than about 8%,not greater than about 7%, not greater than about 6%, not greater thanabout 5%, not greater than about 4%, or not greater than about 3% afterstorage at 2° C.−8° C. (e.g., 4° C.) or 25° C., for about one week, twoweeks, one month, three months, six months, nine months, or twelvemonths, eighteen, or twenty-four months. In an embodiment, the increaseof HMW species is not more than 2%, typically not more than 1%, peryear. By “about” is meant ±20% of a cited numerical value. Thus, about10% means 8% to 12%. Formulations stored as lyophilized productgenerally have less than about 5%, less than about 4%, less than about3%, or less than about 2% HMW species or less than about 5%, less thanabout 4%, less than about 3%, or less than about 2% LMW species afterreconstitution following storage at 2° C.−8° C. (e.g., 4° C.) or 25° C.for about for about one week, two weeks, one month, three months, sixmonths, nine months, or twelve months, eighteen, or twenty-four months.

Formulations of anti-HA antibody molecules can be stored as a liquidfor, e.g., at least two weeks, at least one month, at least two months,at least three months, at least four months, at least six months, atleast nine months, at least one year, or at least two years.Formulations of anti-HA antibody molecules can be stored as a lyophilatefor, e.g., at least two years, at least three years, at least fouryears, or at least five years. In an embodiment, the formulation is in aform of, or is stored as, a frozen lipid.

Additional details related to components of formulations and methods ofassaying the integrity of the anti-HA antibody molecule, e.g., theanti-HA antibody molecule described herein, in a formulation areprovided infra.

Anti-HA antibody molecule concentrations in formulations are generallybetween about 0.1 mg/mL and about 250 mg/mL, e.g., between about 0.5mg/mL and about 100 mg/mL, about 0.5 mg/mL and about 1.0 mg/mL, about0.5 mg/mL and about 45 mg/mL, about 1 mg/mL and about 10 mg/mL, about 5mg/mL and 20 mg/mL, about 8 mg/mL and about 16 mg/mL, about 10 mg/mL andabout 40 mg/mL, about 10 mg/mL and about 50 mg/mL, about 20 mg/mL and 60mg/mL, about 25 mg/mL and 50 mg/mL, about 50 mg/mL and about 100 mg/mL,about 100 mg/mL and about 200 mg/mL, or about 200 mg/mL and about 250mg/mL. In the context of ranges, “about” means −20% of the lower-citednumerical value of the range and +20% of the upper-cited numerical valueof the range. In the context of ranges, e.g., about 10 mg/mL to about100 mg/mL, this means, between 8 mg/mL to 120 mg/mL. In some cases,antibody concentrations in formulations can be, for example, between 1mg/mL and 100 mg/mL, e.g., 2 mg/mL and 80 mg/mL, 5 mg/mL and 60 mg/mL,10 mg/mL and 50 mg/mL, 15 mg/mL and 40 mg/mL, 20 mg/mL and 30 mg/mLanti-HA antibody molecule described herein, e.g., Ab 044. Such antibodyformulations can be used as therapeutic agents. Accordingly, theconcentration of anti-HA antibody molecule in a formulation issufficient to provide such dosages in a volume of the formulation thatis tolerated by a subject being treated and is appropriate for themethod of administration. In one non-limiting example, to supply a highdosage subcutaneously, in which the volume limitation is small (e.g.,about lml to 1.2 ml per injection), the concentration of antibody isgenerally at least 25 mg/mL or greater, e.g., 100 mg/mL or greater,e.g., 100 mg/mL to 500 mg/mL, 100 mg/mL to 250 mg/mL, or 100 mg/mL to150 mg/mL. Such high concentrations can be achieved, for example, byreconstituting a lyophilized formulation in an appropriate volume ofdiluent (e.g., sterile water for injection, buffered saline). In somecases, the reconstituted formulation has a concentration of between 25mg/mL and 500 mg/mL, e.g., between about 100 mg/mL and 500 mg/mL (e.g.,100 mg/mL, 125 mg/mL, 150 mg/mL, 175 mg/mL, 200 mg/mL, 250 mg/mL, 275mg/mL, 300 mg/mL, 350 mg/mL, 375 mg/mL, 400 mg/mL, 425 mg/mL, 450 mg/mL,475 mg/mL and 500 mg/mL). For delivery via inhalation, the formulationis generally somewhat concentrated (e.g., between about 25 mg/mL and 500mg/mL, e.g., between about 100 mg/mL and 500 mg/mL) so as to provide asufficient dose in a limited volume of aerosol for inspiration. In somecases, low concentrations (e.g., between about 0.05 mg/mL and 1 mg/mL)are used. Methods are known in the art to adapt the dosage delivered tothe method of delivery, e.g., a jet nebulizer or a metered aerosol.

Buffers

The pH of a formulation as described herein is generally between aboutpH 5.0 to about 7.0, for example, about pH 5.5 to about 6.5, about pH5.5 to about 6.0, about pH 6.0 to about 6.5, pH 5.5, pH 6.0, or pH 6.5.In general, a buffer that can maintain a solution at pH 5.5 to 6.5 isused to prepare a formulation, e.g., a buffer having a pKa of about 6.0.Suitable buffers include, without limitation, 2-morpholinoethanesulfonicacid (MES), phosphate, and citrate (e.g., citrate-sodium phosphate). Theconcentration of the buffer is between about 5 mM and about 100 mM,e.g., about 25 mM to about 50 mM. In some cases, citrate-sodiumphosphate buffer is used at a concentration of about 40 nM. Otherbuffers can include, histidine buffer, acetate, or succinate, e.g., fora desired pH other than about 6.0, e.g., below 6.0. In other cases,histidine buffer is used at a concentration of up to 60 nM, e.g., about5 mM or about 10 mM. In other cases, acetate or succinate buffer is usedat a concentration of about 5 mM or about 10 mM.

Tonicity Agents

Tonicity agents are known in the art and include, e.g., sodium chloride,potassium chloride, or dextrose.

The tonicity agent is generally used at a concentration of about 50 mMto about 200 mM. For example, the tonicity agent can be used at aconcentration of about 50 mM to about 200 mM, e.g., about 60 mM to about190 mM, about 70 mM to about 180 mM, about 80 mM to about 170 mM, about90 mM to about 160 mM, about 100 mM to about 150 mM, about 145 mM toabout 155 mM, about 140 mM to about 160 mM, about 135 mM to about 165mM, about 130 mM to about 170 mM, about 120 mM to about 180 mM, about110 mM to about 190 mM, about 100 mM to about 200 mM, about 50 mM toabout 100 mM, about 100 mM to about 150 mM, or about 150 mM to about 120mM, e.g., about 200 mM or less, about 150 mM or less, about 100 mM orless, or about 75 mM or less, e.g., about 50 mM, about 60 mM, about 70mM, about 80 mM, about 90 mM, about 100 mM, about 110 mM, about 120 mM,about 130 mM, about 140 mM, about 150 mM, about 160 mM, about 170 mM,about 180 mM, about 190 mM, or about 200 mM.

In an embodiment, the tonicity agent is used at a concentration of about50 to about 200 nM, about 75 mM to about 150 mM, about 120 mM to about180 mM, e.g., about 140 to about 160 mM, e.g., about 150 mM. In anembodiment, the tonicity agent comprises sodium chloride. In anembodiment, the tonicity agent comprises sodium chloride and is used ata concentration of about 140 to about 160 mM, e.g., about 150 mM.

The tonicity agent used in the formulation can generally provide atonicity (or osmolarity) of about 250 mOsm/L to about 350 mOsm/L, about260 mOsm/L to about 340 mOsm/L, about 270 mOsm/L to about 330 mOsm/L,about 280 mOsm/L to about 320 mOsm/L, about 285 mOsm/L to about 310mOsm/L, or about 290 mOsm/L to about 300 mOsm/L, e.g., about 250 mOsm/L,about 260 mOsm/L, about 270 mOsm/L, about 280 mOsm/L, about 290 mOsm/L,about 300 mOsm/L, about 310 mOsm/L, about 320 mOsm/L, about 330 mOsm/L,about 340 mOsm/L, or about 350 mOsm/L.

In an embodiment, the tonicity agent provides a tonicity (or osmolality)of about 240 mOsm/kg to about 340 mOsm/kg, about 250 mOsm/kg to about330 mOsm/kg, about 260 mOsm/kg to about 320 mOsm/kg, about 270 mOsm/kgto about 310 mOsm/kg, about 280 mOsm/kg to about 300 mOsm/kg, or about285 mOsm/kg to about 295 mOsm/kg, e.g., about 240 mOsm/kg, about 250mOsm/kg, about 260 mOsm/kg, about 270 mOsm/kg, about 280 mOsm/kg, about290 mOsm/kg, about 300 mOsm/kg, about 310 mOsm/kg, about 320 mOsm/kg,about 330 mOsm/kg, or about 340 mOsm/kg.

By “isotonic” is meant that the formulation of interest has essentiallythe same osmotic pressure as human blood. Isotonic formulations willgenerally have an osmotic pressure, e.g., from about 250 to 350 mOsm/L.Isotonicity can be measured using a vapor pressure or ice-freezing typeosmometer, for example.

Surfactants

In certain embodiments, a surfactant is included in the formulation.Examples of surfactants include, without limitation, nonionicsurfactants such as polysorbates (e.g., polysorbate-20, polysorbate-40,polysorbate-60, polysorbate-65, polysorbate-80, or polysorbate-85);poloxamers (e.g., poloxamer 188); Triton™; sodium dodecyl sulfate (SDS);sodium laurel sulfate; sodium octyl glycoside; lauryl-sulfobetaine,myristyl-sulfobetaine, linoleyl-sulfobetaine, stearyl-sulfobetaine,lauryl-sarcosine, myristyl-sarcosine, linoleyl-sarcosine,stearyl-sarcosine, linoleyl-betaine, myristyl-betaine, cetyl-betaine,lauroamidopropyl-betaine, cocamidopropyl-betaine,linoleamidopropyl-betaine, myristamidopropyl-betaine,palmidopropyl-betaine, isostearamidopropyl-betaine (e.g.lauroamidopropyl), myristarnidopropyl-, palmidopropyl-, orisostearamidopropyl-dimethylamine; sodium methyl cocoyl-, or disodiummethyl ofeyl-taurate; and the Monaquat™ series (Mona Industries, Inc.,Paterson, N.J.), polyethyl glycol, polyp ropyl glycol, and copolymers ofethylene and propylene glycol (e.g. pluronics, PF68).

The amount of surfactant added is such that it reduces aggregation ofthe reconstituted protein to an acceptable level as assayed using, e.g.,SEC-HPLC of HMW species or LMW species, and minimizes the formation ofparticulates after reconstitution of a lyophilate of an anti-HA antibodymolecule formulation. The addition of surfactant has also been shown toreduce the reconstitution time of a lyophilized formulation of anti-HAantibody molecules, and aid in de-gassing the solution. For example, thesurfactant can be present in the formulation (liquid or prior tolyophilization) in an amount from about 0.001% to 0.5%, e.g., from about0.005% to 0.05%, about 0.005% to about 0.2%, and about 0.01% to 0.2%.

Cryoprotectants

Cryoprotectants are known in the art and include, e.g., sucrose,trehalose, and glycerol. A cryoprotectant exhibiting low toxicity inbiological systems is generally used. The cryoprotectant is included inthe formulation at a concentration of about 0.5% to 15%, about 0.5% to2%, about 2% to 5%, about 5% to 10%, about 10% to 15%, and about 5%(weight/volume).

Histidine buffer, which can be used as a buffer in an anti-HA antibodymolecule formulation, may have cryoprotectant properties. In someembodiments of the invention, a histidine buffer is used in conjunctionwith a cryoprotectant such as a sugar, e.g., sucrose. A formulation ofthe invention can specifically exclude the use of histidine in anysubstantial amount, e.g., neither the buffer nor the cryoprotectantcomponent of the formulation is a histidine.

The viscosity of a formulation is generally one that is compatible withthe route of administration of the formulation. In some embodiments, theviscosity of the formulation is between 1 cP and 2 cP, or similar towater (about 1 cP). In other embodiments, the viscosity of theformulation is between about 5 cP and about 40 cP. In specificembodiments, the viscosity of the formulation is 1 cP, 2 cP, 3 cP, 4 cP,5 cP, 10 cP, 15 cP, 20 cP, 25 cP, 30 cP, 35 cP, or 40 cP.

Additions to Formulations

Formulations are stored as sterile solutions or sterile lyophilates.Prevention of the action of microorganisms in formulations can also beachieved by including at least one antibacterial and/or antifungal agentin a formulation, for example, parabens, chlorobutanol, phenol, ascorbicacid, thimerosal, and the like. In some cases, a lyophilate isreconstituted with bacteriostatic water (e.g., water containing 0.9%benzyl alcohol). Considerations for the inclusion of a preservative in aformulation are known in the art as are methods of identifyingpreservatives that are compatible with a specific formulation and methodof delivery (e.g., see Gupta, et al. (2003), AAPS Pharm. Sci. 5:article8, p. 1-9). A “preservative” is a compound which can be added to thediluent to essentially reduce bacterial action in the reconstitutedformulation, thus facilitating the production of a multi-usereconstituted formulation, for example. Examples of potentialpreservatives include octadecyldimethylbenzyl ammonium chloride,hexamethonium chloride, benzalkonium chloride (a mixture ofalkylbenzyldimethylammonium chlorides in which the alkyl groups arelong-chain compounds), and benzethonium chloride. Other types ofpreservatives include aromatic alcohols such as phenol, butyl and benzylalcohol, alkyl parabens such as methyl or propyl paraben, catechol,resorcinol, cyclohexanol, 3-pentanol, and m-cresol.

In some cases, the formulation is isotonic. In general, any componentknown in the art that contributes to the solution osmolarity/tonicitycan be added to a formulation (e.g., salts, sugars, polyalcohols, or acombination thereof). Isotonicity is generally achieved using either acomponent of a basic formulation (such as sucrose) in an isotonicconcentration or by adding an additional component such as, a sugar, apolyalcohol such as manitol or sorbitol, or a salt such as sodiumchloride.

In some cases, a salt is used in an anti-HA antibody moleculeformulation, e.g., to achieve isotonicity or to increase the integrityof the anti-HA antibody molecule of the formulation. Salts suitable foruse are discussed, supra. The salt concentration can be from 0 mM toabout 300 mM. In one example, the salt is used at a concentration of 150nM in the formulation.

In certain cases, the formulation is prepared with Tween (e.g., Tween®20, Tween® 80) to decrease interfacial degradation. The Tweenconcentration can be from about 0.001% to about 0.05%. In one example,Tween-80 is used at a concentration of 0.025% in the formulation.

In certain other cases, the formulation is prepared with glycine. Theglycine concentration in the formulation can be from about 0.01% toabout 5%. In one example, glycine is used at a concentration of 1% inthe formulation. In another example, glycine is used at a concentrationof 2% in the formulation. In some cases both Tween and arginine areadded to the anti-HA antibody molecule formulations described herein.

In yet other cases, the formulation may be prepared with at least oneof: sucrose, histidine, or arginine. If sucrose is included in theformulation, it can be added to a concentration of between about 1% andabout 10%. In one example, sucrose is found in the formulation at aconcentration of 2%. If histidine is included in the formulation, it canbe added to a concentration of between about 0.5% to about 5%. In oneexample, histidine is found in the formulation at a concentration of 1%.In another example, histidine is found in the formulation at aconcentration of 2%. If arginine is included in the formulation, it canbe added to a concentration of between about 0.5% to about 5%. In oneexample, arginine is found in the formulation at a concentration of 1%.In another example, arginine is found in the formulation at aconcentration of 2%.

Exemplary Formulations

Exemplary anti-HA antibody molecule formulations are described in Table7. In an embodiment, an anti-HA antibody molecule formulation comprises25 mg/mL anti-HA antibody molecule described herein (e.g., Ab 044), 40mM citrate-sodium phosphate, 150 mM sodium chloride, 0.025% Tween-80, atpH 6.0. In another embodiment, an anti-HA antibody molecule formulationcomprises 25 mg/mL anti-HA antibody molecule described herein (e.g., Ab044), 40 mM citrate-sodium phosphate, 150 mM sodium chloride, 0.025%Tween-80, at pH 6.5. In another embodiment, an anti-HA antibody moleculeformulation comprises 25 mg/mL anti-HA antibody molecule describedherein (e.g., Ab 044), 40 mM citrate-sodium phosphate, 1% glycine, 75 mMsodium chloride, 0.025% Tween-80, at pH6.5. In another embodiment, ananti-HA antibody molecule formulation comprises 25 mg/mL anti-HAantibody molecule described herein (e.g., Ab 044), 40 mM citrate-sodiumphosphate, 150 mM sodium chloride, at pH 6.0. In another embodiment, ananti-HA antibody molecule formulation comprises 25 mg/mL anti-HAantibody molecule described herein (e.g., Ab 044), 40 mM citrate-sodiumphosphate, 75 mM sodium chloride, 0.025% Tween-80, at pH 6.5.

Additional exemplary anti-HA antibody molecule formulations aredescribed in Table 5.

Storage and Preparation Methods

Liquid

In some cases, formulations containing antibodies are stored as liquid.Accordingly, it is desirable that the formulation be relatively stableunder such conditions, including, at 4° C. or in room temperature. Onemethod of determining the suitability of a formulation is to subject asample formulation to agitation or storage (e.g., at 4° C., 25° C., or45° C.) for a period of time (e.g., one week, two weeks, or four weeks),determining the amount of LMW species and/or HMW species that accumulateafter the agitation or storage and comparing it to the amount of LMWspecies or HMW species present in the sample prior to the agitation orstorage procedure. An increase in the LMW or HMW species indicatesdecreased stability.

Freezing

In some cases, formulations containing antibodies are frozen forstorage. Accordingly, it is desirable that the formulation be relativelystable under such conditions, including, under freeze-thaw cycles. Onemethod of determining the suitability of a formulation is to subject asample formulation to at least two, e.g., three, four, five, eight, ten,or more cycles of freezing (at, for example −20° C. or -80° C.) andthawing (for example by fast thaw in a 37° C. water bath or slow thaw at2°−8° C.), determining the amount of LMW species and/or HMW species thataccumulate after the freeze-thaw cycles and comparing it to the amountof LMW species or HMW species present in the sample prior to thefreeze-thaw procedure. An increase in the LMW or HMW species indicatesdecreased stability.

Lyophilization Formulations can be stored after lyophilization.Therefore, testing a formulation for the stability of the proteincomponent of the formulation after lyophilization is useful fordetermining the suitability of a formulation. The method is similar tothat described, supra, for freezing, except that the sample formulationis lyophilized instead of frozen, reconstituted to its original volume,and tested for the presence of LMW species and/or HMW species. Thelyophilized sample formulation is compared to a corresponding sampleformulation that was not lyophilized. An increase in LMW or HMW speciesin the lyophilized sample compared to the corresponding sample indicatesdecreased stability in the lyophilized sample.

In general, a lyophilization protocol includes loading a sample into alyophilizer, a pre-cooling period, freezing, vacuum initiation, rampingto the primary drying temperature, primary drying, ramping to thesecondary drying temperature, secondary drying, and stoppering thesample. Additional parameters that can be selected for a lyophilizationprotocol include vacuum (e.g., in microns) and condenser temperature.Suitable ramp rates for temperature are between about 0.1° C./min. to 2°C./min., for example 0.1° C./min. to 1.0° C./min., 0.1° C./min. to 0.5°C./min., 0.2° C./min. to 0.5° C./min., 0.1° C./min., 0.2° C./min., 0.3°C./min., 0.4° C./min., 0.5° C./min., 0.6° C./min., 0.7° C./min., 0.8°C./min., 0.9° C./min., and 1.0° C./min. Suitable shelf temperaturesduring freezing for a lyophilization cycle are generally from about −55°C. to −5° C., −25° C. to −5° C., −20° C. to −5° C., −15° C. to −5° C.,−10° C. to −5° C., −10° C., −11° C., −12° C., −13° C., −14° C., −15° C.,−16° C., −17° C., −18° C., −19° C., −20° C., −21° C., −22° C., −23° C.,−24° C., or −25° C. Shelf temperatures can be different for primarydrying and secondary drying, for example, primary drying can beperformed at a lower temperature than secondary drying. In anon-limiting example, primary drying can be executed at 0° C. andsecondary drying at 25° C.

In some cases, an annealing protocol is used during freezing and priorto vacuum initiation. In such cases, the annealing time must be selectedand the temperature is generally above the glass transition temperatureof the composition. In general, the annealing time is about 2 to 15hours, about 3 to 12 hours, about 2 to 10 hours, about 3 to 5 hours,about 3 to 4 hours, about 2 hours, about 3 hours, about 5 hours, about 8hours, about 10 hours, about 12 hours, or about 15 hours. Thetemperature for annealing is generally from about −35° C. to about −5°C., for example from about −25° C. to about −8° C., about −20° C. toabout −10° C., about −25° C., about −20° C., about −15° C., about 0° C.,or about −5° C. In some cases, the annealing temperature is generallyfrom −35° C. to 5° C., for example from 25° C. to −8° C., −20° C. to−10° C., −25° C., −20° C., −15° C., 0° C., or 5° C.

In general, a lyophilization cycle can run from 10 hours to 100 hours,e.g., 20 hours to 80 hours, 30 hours to 60 hours, 40 hours to 60 hours,45 hours to 50 hours, 50 hours to 65 hours.

Non-limiting examples of the temperature range for storage of anantibody formulation are about −20° C. to about 50° C., e.g., about −15°C. to about 30° C., about −15° C. to about 20° C., about 5° C. to about25° C., about 5° C. to about 20° C., about 5° C. to about 15° C., about2° C. to about 12° C., about 2° C. to about 10 C, about 2° C. to about8° C., about 2° C. to about 6° C., 2 C, 3° C., 4° C., 5° C., 6° C., 7°C., 8° C., 10° C., 15° C., or 25° C. Notwithstanding the storagetemperatures, in certain cases, samples are stable under temperaturechanges that may transiently occur during storage and transportationconditions that can be anticipated for such compositions.

Spray-Drying

In some cases, a formulation is spray-dried and then stored.Spray-drying is conducted using methods known in the art, and can bemodified to use liquid or frozen spray-drying (e.g., using methods suchas those from Niro Inc. (Madison, Wis.), Upperton Particle Technologies(Nottingham, England), or Buchi (Brinkman Instruments Inc., Westbury,N.Y.), or U.S. Application Pulication Nos. 2003/0072718 and2003/0082276).

Determination of Antibody Molecule Integrity

The accumulation of LMW species and HMW species are useful measures ofantibody stability. Accumulation of either LMW or HMW in a formulationis indicative of instability of a protein stored as part of theformulation. Size exclusion chromatography with HPLC can be used todetermine the presence of LMW and HMW species. Suitable systems for suchmeasurements are known in the art, e.g., HPLC systems (Waters, Milford,Mass.). Other systems known in the art can be used to evaluate theintegrity of antibody in a formulation, for example, SDS-PAGE (tomonitor HMW and LMW species), bioassays of antibody activity,enzyme-linked immunosorbent assay, ability to bind purified targetprotein (e.g., HA), and cation exchange-HPLC (CEX-HPLC; to detectvariants and monitor surface charge). In one example, a bioassay is acell-based assay in which inhibition of an HA-dependent activity isexamined in the presence of different concentrations of formulatednanobody molecule to demonstrate biological activity.

Articles of Manufacture

The present application also provides an article of manufacture thatincludes a formulation as described herein and provides instructions foruse of the formulation. The article of manufacture can include acontainer suitable for containing the formulation. A suitable containercan be, without limitation, a bottle, vial, syringe, test tube,nebulizer (e.g., ultrasonic or vibrating mesh nebulizers), i.v. solutionbag, or inhaler (e.g., a metered dose inhaler (MDI) or dry powderinhaler (DPI)). The container can be formed of any suitable materialsuch as glass, metal, or a plastic such as polycarbonate, polystyrene,or polypropylene. In general, the container is of a material that doesnot absorb significant amounts of protein from the formulation and isnot reactive with components of the formulation. In some embodiments,the container is a clear glass vial with a West 4432/50 1319 siliconizedgray stopper or a West 4023 Durafluor stopper. In some embodiments, thecontainer is a syringe. In specific embodiments, the formulationcomprises about 25 mg/mL of an antibody molecule described herein, about40 mM citrate-sodium phosphate, about 150 mM sodium chloride, and about0.025% polysorbate 80, at a pH of about 6, in a pre-filled syringe. Incertain embodiments, the syringe is suitable for use with anauto-injector device.

In an embodiment, the container is a container suitable for storage ofthe formulation or antibody molecule, e.g., a vial. In anotherembodiment, the container is a container suitable for administration ofthe formulation or antibody molecule, e.g., an intravenous (IV) bag. Inan embodiment, the antibody molecule or formulation in a first container(e.g., suitable for storage) is transferred to a second container (e.g.,suitable for administration) before use. In an embodiment, transferincludes dilution of the antibody molecule or formulation. In anembodiment, transfer occurs less than 4 hours, e.g., less than 3, 2, or1 hours, prior to administration of the antibody molecule or formulationto a subject.

In an embodiment, the container suitable for administration (e.g., an IVsolution bag) is a primary container and ready to use for administration(e.g., IV administration). For example, in one configuration, it istypically not necessary, or there is no need, to transfer the antibodymolecule or formulation, e.g., from a vial (e.g., a storage vial) to anIV solution bag, or to dilute the antibody molecule or formulation,e.g., into an IV solution, before administration (e.g., on the same dayof administration). In an embodiment, the container is a vial, e.g., aglass vial. In an embodiment, the container (e.g., vial) comprises about10 mg/mL to about 100 mg/mL, e.g., about 20 mg/mL to about 60 mg/mL(e.g., about 25 mg/mL to about 50 mg/mL) of the antibody molecule. In anembodiment, the container (e.g., vial) comprises about 10 mL to about 60mL, e.g., about 20 mL to about 40 mL, of the antibody molecule orformulation. In an embodiment, the container (e.g., vial) is a first (orprimary) container, e.g., for storing the antibody molecule orformulation.

The antibody molecule or formulation can be transferred into a secondcontainer before use. In an embodiment, the second container issuitable, or includes a solution that is suitable, for administration,e.g., intravenous administration. In an embodiment, the second containerincludes a solution suitable for intravenous administration. In anembodiment, the solution comprises saline, optionally, further comprisesdextrose. In an embodiment, the solution (e.g., saline) does notcomprise dextrose. For example, an amount equal to one dose of theantibody molecule can be transferred into a container suitable for IVadministration. In an embodiment, 1 to 10 vials (e.g., 1 to 8 vials, 1to 6 vials, 1 to 4 vials, 1 to 2 vials, 6 to 8 vials, 4 to 8 vials, or 2to 8 vials) of the antibody molecule or formulation are diluted into anIV solution bag, e.g., containing saline with or without dextrose.

In an embodiment, the container is a container suitable for IVadministration (e.g., an IV solution bag). In an embodiment, the amountof the antibody molecule in the container (e.g., IV solution bag) equalsto 1 to 10 vials (e.g., 1 to 8 vials, 1 to 6 vials, 1 to 4 vials, 1 to 2vials, 6 to 8 vials, 4 to 8 vials, or 2 to 8 vials) of the antibodymolecule as described above. In an embodiment, the container (e.g., IVsolution bag) comprises about 500 mg to about 16000 mg, e.g., about 500mg to about 8000 mg, about 500 mg to about 5000 mg/mL, about 1000 mg toabout 5000 mg, about 2000 mg to about 4000 mg, or about 2300 mg to about4600 mg, e.g., about 2300 mg or about 4600 mg, of the antibody moleculeor formulation. In an embodiment, the container (e.g., IV solution bag)further comprises saline. In an embodiment, the container furthercomprises dextrose. In another embodiment, the container does notcomprise dextrose.

In an embodiment, the container suitable for IV administration (e.g., IVsolution bag) is not a second (or secondary) container (e.g., is a first(or primary) container, e.g., where the antibody molecule is stored),and comprises about 5 mg/mL to about 25 mg/mL, e.g., about 8 mg/mL toabout 16 mg/mL of the antibody molecule. In an embodiment, the container(e.g., IV solution bag) comprises about 100 mL to about 400 mL, e.g.,about 200 mL to about 300 mL, of antibody molecule. In an embodiment,the container (e.g., IV solution bag) comprises about 2000 mg to about5000 mg, e.g., about 2300 mg to about 4600 mg, of the antibody molecule.

In an embodiment, the antibody molecule is administered from thecontainer (e.g., IV solution bag) to the subject through an IV line.

Disclosed herein are also methods of preparing a composition (e.g., asolution) or a container for administration (e.g., intravenousadministration). In an embodiment, the method comprises transferring anantibody molecule or a formulation disclosed herein to a containersuitable for administration (e.g., an intravenous (IV) solution bag). Inan embodiment, the method comprises contacting, e.g., combining (e.g.,mixing or diluting) an antibody molecule or a formulation disclosedherein with a solution suitable for administration. In an embodiment,the container suitable for administration is an IV solution bag. In anembodiment, the solution suitable for administration is an IV solution,e.g., saline with or without dextrose. In an embodiment, about 2000 mgto about 5000 mg of the antibody molecule is contacted (e.g., combined)with the solution. In an embodiment, about 2300 mg to about 4600 mg orabout 2000 mg to about 4000 mg of the antibody molecule is contacted(e.g., combined) with the solution.

Examples of nebulizers include, in non-limiting examples, jetnebulizers, ultrasonic nebulizers, and vibrating mesh nebulizers. Theseclasses use different methods to create an aerosol from a liquid. Ingeneral, any aerosol-generating device that can maintain the integrityof the protein in these formulations is suitable for delivery offormulations as described herein.

Formulations to be used for administration to a subject, e.g., as apharmaceutical, must be sterile. This is accomplished using methodsknown in the art, e.g., by filtration through sterile filtrationmembranes, prior to, or following, formulation of a liquid orlyophilization and reconstitution. Alternatively, when it will notdamage structure, components of the formulation can be sterilized byautoclaving and then combined with filter or radiation sterilizedcomponents to produce the formulation.

Hemagglutinin (HA) Polypeptides and Influenza

Influenza viruses are negative sense, single-stranded, segmented RNAenvelope viruses. Two glycoproteins, a hemagglutinin (HA) polypeptideand a neuraminidase (NA) polypeptide, are displayed on the outer surfaceof the viral envelope. There are several Influenza A subtypes, labeledaccording to an H number (for the type of hemagglutinin) and an N number(for the type of neuraminidase). There are 17 different H antigens (H1to H17) and nine different N antigens (N1 to N9). Influenza strains areidentified by a nomenclature based on the number of the strain's HApolypeptide and NA polypeptide subtypes, for example, H1N1, H1N2, H1N3,H1N4, H1N5, and the like.

HA is the major viral surface glycoprotein that mediates binding andentry of the virus into host cells and is a primary target ofneutralizing antibody responses. HA is a trimer of three identicalmonomers. Each monomer is synthesized as a precursor, HA₀, that isproteolytically processed into two disulfide-bonded polypeptide chains,HA₁ and HA₂. The ectodomain of this protein has (i) a globular headdomain possessing receptor binding activity and major antigenicdeterminants, (ii) a hinge region, and (iii) a stem region where asequence critical for fusion, the fusion peptide, is located. The viralreplication cycle is initiated when the virion attaches via its surfacehemagglutinin proteins to sialylated glycan receptors on the host celland enters the cell by endocytosis. The acidic environment in theendosome induces conformational changes in HA that expose the fusionpeptide hidden within the stem region of the trimer. The exposed fusionpeptide mediates the fusion of the viral and target cell membranesresulting in the release of the viral ribonucleoprotein into the cellcytoplasm.

Influenza A hemagglutinin subtypes have been divided into two maingroups and four smaller clades, and these are further divided intoclusters. Group 1 influenza A strains are divided into 3 clades: (i) H8,H9 and H12 (“the H9 cluster”); (ii) H1, H2, H5, H6 and H17 (“the H1acluster”); and (iii) H11, H13 and H16 (“the H1b cluster”). Group 2strains are divided into 2 clades: (i) H3, H4 and H14 (“the H3cluster”); and (ii) H7, H10 and H15 (“the H7 cluster”). The H1b and theH1a clusters are classified together as the H1 cluster. The different HAsubtypes do not necessarily share strong amino acid sequence identity,but their overall 3D structures are similar.

Of the 17 HA polypeptide subtypes, only 3 (H1, H2 and H3) have adaptedfor human infection. These subtypes have in common an ability to bindalpha 2,6 sialylated glycans. In contrast, their avian counterpartspreferentially bind to alpha 2,3 sialylated glycans. HA polypeptidesthat have adapted to infect humans (e.g., of HA polypeptides from thepandemic H1N1 (1918) and H3N2 (1967-68) influenza subtypes) have beencharacterized by an ability to preferentially bind to α2,6 sialylatedglycans in comparison with their avian progenitors that preferentiallybind to α2,3 sialylated glycans (see e.g., Skehel & Wiley, Annu RevBiochem, 69:531, 2000; Rogers, & Paulson, Virology, 127:361, 1983;Rogers et al., Nature, 304:76, 1983; Sauter et al., Biochemistry,31:9609, 1992).

Further, HA polypeptides that mediate infection of humans preferentiallybind to umbrella topology glycans over cone topology glycans (see e.g.,U.S. 2011/0201547). Without wishing to be bound by any particulartheory, it has been proposed that the ability to infect human hostscorrelates less with binding to glycans of a particular linkage, andmore with binding to glycans of a particular topology, even thoughcone-topology glycans may be α2,6 sialylated glycans. In has beendemonstrated that HA polypeptides that mediate infection of humans bindto umbrella topology glycans, often showing preference for umbrellatopology glycans over cone topology glycans (see, for example, U.S. Ser.No. 12/348,266 filed Jan. 2, 2009, U.S. Ser. No. 12/301,126, filed Nov.17, 2008, U.S. Ser. No. 61/018,783, filed Jan. 3, 2008, U.S. Ser. No.11/969,040, filed Jan. 3, 2008, U.S. Ser. No. 11/893,171, filed Aug. 14,2007, U.S. Ser. No. 60/837,868, filed on Aug. 14, 2006, U.S. Ser. No.60/837,869, filed on August 14, and to PCT application PCT/US07/18160,filed Aug. 14, 2007).

Mature HA polypeptides include three domains, (i) a globular domain(a.k.a., the head domain) consists mainly of the HA1 peptide andcontains the receptor (sialylated glycoproteins)-binding region, (ii) astalk domain (HA1 and HA2) where the membrane fusion peptide resides,and (iii) a transmembrane domain (HA2) that anchors hemagglutinin to theviral envelope. A set of amino acids in the interface of the HA1 and HA2peptides is highly conserved across all influenza subtypes. The HA1/HA2membrane proximal region (MPER), including a canonical alpha-helix, isalso highly conserved across influenza subtypes.

HA polypeptides interact with the surface of cells by binding to aglycoprotein receptor, known as the HA receptor. Binding of an HApolypeptide to an HA receptor is predominantly mediated by N-linkedglycans on the HA receptors. HA polypeptides on the surface of flu virusparticles recognize sialylated glycans that are associated with HAreceptors on the surface of the cellular host. Following replication ofviral proteins and genome by the cellular machinery, new viral particlesbud from the host to infect neighboring cells.

Currently, vaccines are administered to subjects, e.g., humans, toprevent the flu, e.g., to prevent infection or to minimize the effectsof an infection with influenza virus. Traditional vaccines contain acocktail of antigens from various strains of influenza and areadministered to humans to prevent the human from getting infected withthe virus. HA is the main target of influenza A-neutralizing antibodies,and HA undergoes continuous evolution driven by the selective pressureof the antibody response, which is primarily directed against themembrane-distal receptor-binding subdomain of the HA polypeptide. Thesubject, however, is protected only from strains that are identical to,or closely related to, the strains from which the antigens in thecocktail were derived. The human is still most vulnerable to infectionby other strains of the flu that were not included in the cocktail. Oneof the advantages of the antibodies provided herein is their ability tobind an epitope of HA that is conserved across multiple strains ofinfluenza A, and in an embodiment, influenza B. Thus, administration ofan anti-HA antibody described herein will be more effective to protectan individual from infection from a broader spectrum of influenza (e.g.,influenza A and, in an embodiment, influenza B) and conditions associatethereof (e.g., secondary infections, e.g., secondary bacterialinfections). Further, the antibodies are effective in treating a subjectafter infection has occurred.

Epitope

HAs exist in nature as homotrimers of proteolytically processed maturesubunits. Each subunit of the trimer is synthesized as a precursor. Aprecursor molecule is proteolytically processed into two disulfidebonded polypeptide chains to form a mature HA polypeptide. The mature HApolypeptide includes two domains: (1) a core HA-1 domain that extendsfrom the base of the molecule through the fibrous stem to the membranedistal head region that contains the glycan receptor binding domain,returning to fibrous region ending in the cleavage site, and (2) HA-2domain that includes the stem region and the transmembrane domain of HA.HA-1 includes a glycan binding site. The glycan binding site may beresponsible for mediating binding of HA to the HA-receptor. The HA-2domain acts to present the HA-1 domain. The HA trimer can be stabilizedby polar and non-polar interactions between the three long HAalpha-helices of the stem of HA monomers.

HA sequences from all influenza subtypes share a set of amino acids inthe interface of the HA-1 and HA-2 domains that are well conserved. TheHA-1/HA-2 interface membrane proximal epitope region (MPER) thatincludes the canonical α-helix and residues in its vicinity are alsoconserved across a broad spectrum of subtypes. (Ekiert et al., Science.324(5924):246, 2009; Sui et al., Nat Struct Mol Biol. 16(3):265, 2009).

Ab 044 has high affinity for HA's from Group 1 and Group 2. It binds aconformational epitope that is broadly conserved across a plurality ofinfluenza strains. Numerous amino acid residues distributed along thelinear sequences of HA from different strains/subtypes contribute the Ab044 conformational epitope. The interaction of Ab044 with H3 wasanalyzed by docking studies and residues bound by (or not bound by) Ab044 were identified.

The Fv of Ab 044 was docked against HA of group I and II strains usingZDOCK. The structure of the HA antigen was modeled using the SWISS MODELhomology modeling server keeping the solved crystal structure of H1N1 asthe template. ZDOCK uses shape complementarity along with desolvationand electrostatic energy terms (‘ZRANK’) to rank docked poses. To ensurethe docked poses do not deviate significantly from the native complex,mapped epitope and paratope residues by alanine scanning are forced tobe included in the binding interface.

For comparison studies, amino acids that bind (or do not bind) FI6 weretaken from published US patent application US 2011/0274702 A1,Neutralizing Anti-Influenza A Virus Antibodies and Uses Thereof, filedJul. 18, 2011.

ZDOCK is a Fast Fourier Transform based protein docking program. It wasdeveloped by Zhiping Weng at the University of Massachusetts MedicalSchool. In ZDOCK, two PDB files are input and the output is thepredicted structure of their complex. The program searches all possiblebinding modes in the translational and rotational space between the twoproteins and evaluates each by an energy scoring function. The protein'sstructure is converted to a digital signal and a Fast Fourier Transformtechnique used to reduce computational time. ZDOCK is discussed inPierce B G, Hourai Y, Weng Z. (2011) Accelerating Protein Docking inZDOCK Using an Advanced 3D Convolution Library. PLoS One 6(9): e24657,Pierce B, Tong W, Weng Z. (2005) M-ZDOCK: A Grid-based Approach forC_(n) Symmetric Multimer Docking. Bioinformatics 21(8): 1472-1476;Mintseris J, Pierce B, Wiehe K, Anderson R, Chen R, Weng Z. (2007)Integrating Statistical Pair Potentials into Protein Complex Prediction.Proteins 69(3): 511-520; and Chen R, Li L, Weng Z. (2003) ZDOCK: AnInitial-stage Protein Docking Algorithm. Proteins 52(1): 80-7.

SWISS-MODEL is a fully automated protein structure homology-modelingserver. It is accessible via the ExPASy web server, or from the programDeepView (Swiss Pdb-Viewer). Swiss-Model is discussed in Arnold K.,Bordoli L., Kopp J., and Schwede T. (2006). The SWISS-MODEL Workspace: Aweb-based environment for protein structure homology modelling.Bioinformatics, 22,195-201; Kiefer F, Arnold K, Kunzli M, Bordoli L,Schwede T (2009). The SWISS-MODEL Repository and associated resources.Nucleic Acids Research. 37, D387-D392; and Peitsch, M. C. (1995) Proteinmodeling by E-mail Bio/Technology 13: 658-660.

H3 residues that bind Ab 044 and H3 residues that bind FI6 are discussedbelow.

H3 HA1

The amino acid sequence of H3 HA1 is provided below, as SEQ ID NO: 173.Residues N38, I278, and D291 shown in dashed boxes, are bound by Ab 044but not by FI6; Residues Q327, T328, and R329 shown in dotted boxes, arebound by FI6 but not by Ab 044; residues T318, R321, and V323 shown insolid boxes, are bound by both Ab 044 and FI6.

(SEQ ID NO: 173)

IDNNPHRILD GIDCTLIDAL LGDPHCDVFQ NETWDLFVER SKAFSNCYPY DVPDYASLRSLVASSGTLEF ITEGFTWTGV TQNGGSNACK RGPGSGFFSR LNWLTKSGST YPVLNVTMPNNDNFDKLYIW GIHHPSTNQE QTSLYVQASG RVTVSTRRSQ QTIIPNIGSR PWVRGLSSRI

H3 HA2

The amino acid sequence of H3 HA21 is provided below, as SEQ ID NO: 174Residue N12 shown in a dash box, is bound by Ab 044 but not by FI6;Residues G1, L2, F3, G4, and D46 shown in dotted boxes, are bound by FI6but not by Ab 044; residues A7, E11, I18, D19, G20, W21, L38, K39, T41,Q42, A43, I45, I48, N49, L52, N53, I56, and E57, shown in solid boxes,are bound by both Ab 044 and FI6.

(SEQ ID NO: 174)

TDSEMNKLFE KTRRQLRENA EEMGNGCFKI YHKCDNACIE SIRNGTYDHD VYRDEALNNR FQIKG

H1 residues that bind Ab 044 and H1 residues that bind FI6 are discussedbelow.

H1 HA1

The amino acid sequence of H1 HA1 is provided below, as SEQ ID NO: 181.Residues H31, N279, and 5292 shown in dashed boxes, are bound by Ab 044but not by FI6. Residues Q328 and S329 shown in dotted boxes, are boundby FI6 but not by Ab 044. Residues T319, R322, and 1324 shown in solidboxes, are bound by both Ab 044 and FI6.

(SEQ ID NO: 181)

EDSHNGKLCK LKGIAPLQLG KCNIAGWLLG NPECDLLLTA SSWSYIVETSNSENGYCYPG DFIDYEELRE QLSSVSSFEK FEIFPKTSSW PNHETTKGVTAACSYAGASS FYRNLLWLTK KGSSYPKLSK SYVNNKGKEV LVLWGVHHPPTGTDQQSLYQ NADAYVSVGS SKYNRRFTPE IAARPKVRDQ AGRMNYYWTL

H1 HA2

The amino acid sequence of H1 HA2 is provided below, as SEQ ID NO: 182.Residues G12 shown in a dashed box, is bound by Ab 044 but not by FI6.Residues G1, L2, F3, G4, and D46 shown in dotted boxes, are bound by FI6but not by Ab 044. Residues A7, E11, I18, D19, G20, W21, Q38, K39, T41,Q42, N43, 145, 148, T49, V52, N53, 156, and E57 shown in solid boxes,are bound by both Ab 044 and FI6.

(SEQ ID NO: 182)

VLLENERTLD FHDSNVRNLY EKVKSQLKNN AKEIGNGCFE FYHKCDDACMESVRNGTYDY PKYSEESKLN REEIDGVKLE SMGVYQILAI YSTVASSLVLLVSLGAISFW MCSNGSLQCR ICI

A three dimensional representation of H3 HA with the amino acidsresidues that are predicted to be part of Ab044 epitope but not part ofFI6's epitope highlighted (that is, the highlighted amino acids areunique to Ab044's epitope) is shown in FIG. 26 of InternationalApplication Publication No. WO2013/170139. A three dimensionalrepresentation of H3 HA with the amino acid residues that are part ofFI6's epitope but not predicted to be part of Ab044's epitopehighlighted is shown in FIG. 27 of International Application PublicationNo. WO2013/170139. The content of International Application PublicationNo. WO2013/170139 is incorporated by reference in its entirety.

Binding Agents, e.g., Anti-HA Antibody Molecules

Formulations (e.g., pharmaceutical formulations) described hereininclude binding agents, e.g., antibody molecules, described herein.

Binding agents, and in particular, the antibody molecules describedherein, can bind to influenza A viruses from both Group 1 and Group 2,and in an embodiment also bind influenza B viruses. For example, theantibody molecules described herein can bind to an HA polypeptide on atleast 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11 strains from Group 1, and canalso bind to an HA polypeptide on at least 1, 2, 3, 4, 5, or 6 strainsfrom Group 2. In another example, the antibody molecules describedherein can bind to an HA polypeptide on an influenza strain from atleast 1, 2 or 3 clades from Group 1, and can also bind to an HApolypeptide on an influenza strain from one or both clades of Group 2.The antibody molecules described herein inhibit cell entry and thustargeting an early step in the infection process.

The binding agents, and in particular, the antibody molecules disclosedherein, can be effective to treat or prevent infection by seasonal orpandemic influenza strains. The binding agents, and in particular theantibody molecules described herein, can be characterized by theirability to prevent or treat a Group 1 or a Group 2 strain of influenza Aviruses or, in an embodiment, a strain of influenza B viruses. Thebinding agents, and in particular the antibody molecules disclosedherein, are effective to prevent or treat infection by one or morestrains of Group 1, one or more strains of Group 2, and also one or morestrains of influenza B viruses.

The binding agents, and in particular the antibody molecules can beeffective to treat the infection when administered the same day as thesubject is exposed, or when administered, e.g., 1 day, 2 days, 3 days, 4days or later after infection, or upon a first symptom experienced bythe patient.

Strains

The antibody molecules described herein are effective to treat one ormore influenza strains of Group 1, one or more influenza strains ofGroup 2, and also one or more influenza B strains, and specific isolateswithin these strains. Certain antibody molecules may be more effectivefor treatment of certain isolates than other isolates. Exemplaryinfluenza strains and isolates are described in the below Table 1.

TABLE 1 Exemplary influenza strains and Isolates Type Group HA typeIsolate A 1 H1N1 A/PR/8/34 (aka PR-8) A/Solomon Islands/03/06 A/SolomonIslands/20/1999 A/California/07/2009 A/New Caledonia/20/99A/Bangkok/10/83 A/Yamagata/120/86 A/Osaka/930/88 A/Suita/1/89A/California/04/2009 A 1 H2N2 A/Okuda/57 A/Adachi/2/57 A/Kumamoto/1/65A/Kaizuka/2/65 A/Izumi/5/65 A/Chicken/PA/2004 A 1 H5N1 A/Vietnam/1203/04A/Duck/Singapore/3/97 A/Duck/MN/1525/81 A 1 H9N2 A/Hong Kong/1073/2004A/Swine/Hong Kong/9/98 A/Guinea fowl/HK/WF10/99 A 1 H16N3 A/black headedgull/ Mongolia/1756/2006 A 2 H3N2 X-31 A/Victoria/3/75 A/Wyoming/03/2003A/Wisconsin/67/2005 A/Brisbane/10/2007 A/California/7/2004 A/NewYork/55/2004 A/Moscow/10/1999 A/Aichi/2/68 A/Beijing/32/92/X-117A/Fukuoka/C29/85 A/Sichuan/2/87 A/Ibaraki/1/90 A/Suita/1/90A/Perth/16/2009 A/Uruguay/716/2007 A/Fujian/411/2003 A/Panama/2007/99A/Shangdong/09/93 A 2 H7N7 A/Netherlands/219/2003 A 2 H7N9A/Anhui/1/2013 A/Shanghai/1/2013 B B/Wisconsin/1/2010

Affinity can also be in reference to a particular isolate of a givenGroup 1 or Group 2 strain for influenza A viruses or a strain forinfluenza B viruses. Exemplary isolates are as provided in the aboveTable 1.

Mechanisms of Inhibition

While not being limited by a specific mechanism, HA specific antibodiescan inhibit infection by numerous methods, such as by blocking viralattachment to sialic acid residues on surface proteins on host cells, byinterfering with the structural transition of HA that triggers fusionactivity in the endosome, or by simultaneously inhibiting attachment andvirus-cell fusion.

In an embodiment, antibody molecules disclosed herein bind an epitope atthe HA trimer interface. Structural changes at the trimer interface areimportant for fusion of the viral membrane and the endocytic membrane,and the antibody molecules described herein interfere with this criticalstep of infection. Assays to measure fusogenic activity of HA are knownin the art. For example, one fusion assay measures syncytia formation,which occurs in cell-cell fusion events. Cells that express and displayan influenza viral strain HA can be used in the assay. Membrane-anchoredhemagglutinin in these cells is induced to convert to the fusionconformation by a brief (e.g., 3 minute) exposure to low pH (e.g., pH5). A 2-3-hour incubation period follows to allow the cells to recoverand fuse to form syncytia. A nuclear stain can be used to aid in thevisualization of these fusion products, and their count is used as agauge of fusion activity. A candidate anti-HA antibody can be addedeither before or after the low pH treatment to determine at which stageof the fusion process the antibody interferes.

Another type of fusion assay monitors content mixing. To measure contentmixing, host cells (e.g., erythrocytes) are loaded with a dye (e.g.,Lucifer yellow) to determine whether the contents of HA-bound host cellscould be delivered to HA-expressing cells after exposure tofusion-inducing conditions (e.g., low pH, such as pH less than 6 or pHless than 5). If the dye fails to mix with the contents of the hostcells, then the conclusion can be made that fusion is inhibited. Seee.g., Kemble et al., J. Virol. 66:4940-4950, 1992.

In another example, a fusion assay is performed by monitoring lipidmixing. The lipid mixing assay can be performed by labeling host cells(e.g., erythrocytes) with a fluorescent dye (e.g., R18(octadecylrhodamine)) or dye pairs (e.g., CPT-PC/DABS-PC) (forfluorescence resonance energy transfer), exposing the host cells andHA-expressing cells to fusion-inducing conditions, and assaying forfluorescence dequenching (FDQ). Lipid mixing leads to dilution of thelabel into the viral envelope and a consequent dequenching. A lag indequenching or the absence of dequenching is indicative of membranefusion inhibition. See e.g., Kemble et al., J. Virol. 66:4940-4950,1992; and Carr et al., Proc. Natl. Acad. Sci. 94:14306-14313, 1997.

Escape Mutants

In an embodiment, influenza strains will rarely if ever produce escapemutants when contacted with the formulations (e.g., pharmaceuticalformulations) described herein.

Escape mutants can be identified by methods known in the art. Forexample, a formulation (e.g., pharmaceutical formulation) will notproduce an escape mutant when the cells are infected with the virusunder prolonged or repeated exposure to the formulation (e.g.,pharmaceutical formulation).

One exemplary method includes infection of cells (e.g., MDCK cells) witha fixed amount of influenza A viral particles in the presence of theantibody at a concentration known to attenuate infection rates by 50%.Viral progeny collected after each passaging is used to infect a freshcell culture in the presence of the same or greater concentration of theantibody. After multiple cycles of infection, e.g., after 15 cycles, 12cycles, 11 cycles, 10 cycles, 9 cycles, 8 cycles, 7 cycles, 6 cycles, or5 cycles, of infection under these conditions, the HA nucleotidesequence extracted from 20 viral plaque picks is evaluated forenrichment for mutations that renders the viral isolate resistant toneutralization by the antibody (an escape mutant). If no mutants withreduced sensitivity to the antibody are detected after the multiplerounds of selection, e.g., after 11 rounds, 10 rounds, or 9 rounds ofselection, the antibody is determined to be resistant to escapemutations (see e.g., Throsby et al. (2008) PLoS One, volume 3, e3942).

In another example, an assay that measures minimum inhibitoryconcentration (MIC) of the neutralizing antibody can be used to identifyescape mutants. The MIC of an antibody molecule is the lowestconcentration of an antibody molecule that can be mixed with virus toprevent infection of cell culture with influenza. If escape mutantsarise within a viral population, then the MIC of a particular antibodywill be observed to increase with increased rounds of propagation underthe antibody selective pressure, as the proportion of the viralparticles that carry the resistance mutation within the populationincreased. Influenza escape mutants rarely if ever evolve in response toan anti-HA antibody molecule described herein, and therefore the MICwill stay the same over time.

Another assay suitable for monitoring for the development of escapemutants is a Cytopathic Effect (CPE) assay. A CPE assay monitors theability of an antibody to neutralize (e.g., prevent infection by) aninfluenza strain. A CPE assay provides the minimal concentration ofantibody required in cell culture to neutralize the virus. If escapemutants arise, than the CPE of a particular antibody will increase overtime, as the antibody becomes less effective at neutralizing the virus.Viral strains rarely if ever produce escape mutants in response to ananti-HA antibody molecule described herein, and therefore the CPE willstay essentially the same over time.

Quantitative polymerase chain reaction (qPCR) can also be used tomonitor for the development of escape mutants. qPCR is useful to monitorthe ability of an antibody to neutralize (e.g., prevent infection by) aninfluenza strain. If an antibody effectively neutralizes a virus, thenqPCR performed on cell culture samples will not detect presence of viralgenomic nucleic acid. If escape mutants arise, than over time, qPCR willamplify more and more viral genomic nucleic acid. Escape mutants rarelyif ever develop in response to an anti-HA antibody molecule describedherein, and therefore qPCR will rarely if ever detect viral genomicnucleic acid, even after the passage of time.

Binding and Affinity

In an embodiment, the binding agents, particularly antibody molecules,described herein bind to two or more of the following: at least one HApolypeptide from a Group 1 influenza strain (e.g., an H1, H2, H5, H6,H8, H9 H12, H11, H13, H16 or H17 polypeptide); at least one HApolypeptide from a Group 2 influenza strain (e.g., an H3, H4, H14, H7,H10, or H15 polypeptide); and at least one HA polypeptide from ainfluenza B strain.

In an embodiment, a binding agent, e.g., an antibody molecule, has aK_(D) for an HA from a Group 1 influenza strain (e.g., an H1, H2, H5,H6, H8, H9 H12, H11, H13, H16 or H17 polypeptide) of equal to or lessthan 10⁻⁶, 10⁻⁷, 10⁻⁸, 10⁻⁹, 10⁻¹⁰, 10⁻¹¹, or 10⁻¹² nM. In anembodiment, a binding agent, e.g., an antibody molecule, has a K_(D) foran HA from a Group 2 influenza strain (e.g., an H3, H4, H14, H7, H10, orH15 polypeptide) of equal to or less than 10⁻⁶, 10⁻⁷, 10⁻⁸, 10⁻⁹, 10⁻¹⁰,10⁻¹¹, or 10⁻¹² nM. In an embodiment, a binding agent, e.g., an antibodymolecule, has a K_(D) for an influenza B HA of equal to or less than10⁻⁶, 10⁻⁷, 10⁻⁸, 10⁻⁹, 10⁻¹⁰, 10⁻¹¹, or 10⁻¹² nM.

In an embodiment, a binding agent, e.g., an antibody molecule, has: a) afirst K_(D) (representing an affinity for an HA from a Group 1 influenzastrain, e.g., an H1, H2, H5, H6, H8, H9 H12, H11, H13, H16 or H17polypeptide); and b) a second K_(D) (representing an affinity for an HAfrom a Group 2 influenza strain, e.g., an H3, H4, H14, H7, H10, or H15polypeptide), wherein the first and second K_(D) are one or both of:both equal to or less than 10⁻⁸ nM; and within 10 or 100 fold of eachother.

In an embodiment, a binding agent, e.g., an antibody molecule, has: a) afirst K_(D) (representing an affinity for an H1, e.g., the H1 from anH1N1 strain, e.g., A/South Carolina/1/1918, A/Puerto Rico/08/1934, orA/California/04/2009, or an H5N1 strain, e.g., A/Indonesia/5/2005 orA/Vietnam/1203/2004); and b) a second K_(D) (representing an affinityfor an H3 polypeptide, e.g., the H3 from an H3N2 strain, e.g.,A/Brisbane/59/2007), wherein the first and second K_(D) are one or bothof: both equal to or less than 10⁻⁸ nM; and within 10 or 100 fold ofeach other.

In an embodiment, a binding agent, e.g., an antibody molecule, has: a) afirst K_(D) (representing an affinity for an H1, e.g., the H1 from anH1N1 strain, e.g., A/South Carolina/1/1918, A/Puerto Rico/08/1934, orA/California/04/2009, or an H5N1 strain, e.g., A/Indonesia/5/2005 orA/Vietnam/1203/2004); and b) a second K_(D) (representing an affinityfor an H3 polypeptide, e.g., the H3 from an H3N2 strain, e.g.,A/Brisbane/59/2007), wherein the first and second K_(D) are one or bothof: both equal to or less than 10⁻⁸ nM; and within 10 or 100 fold ofeach other.

In an embodiment, a binding agent, e.g., an antibody molecule, has: a) afirst K_(D) (representing an affinity for an HA from a Group 1 influenzastrain, e.g., an H1, H2, H5, H6, H8, H9 H12, H11, H13, H16 or H17polypeptide and/or an affinity for an HA from a Group 2 influenzastrain, e.g., an H3, H4, H14, H7, H10, or H15 polypeptide); and b) asecond K_(D) (representing an affinity for an influenza B HA, e.g., fromB/Wisconsin/1/2010), wherein the first and second K_(D) are one or bothof: both equal to or less than 10⁻⁸ nM; and within 10 or 100 fold ofeach other.

In an embodiment, a binding agent, e.g., an antibody molecule, has: a) afirst K_(D) (representing an affinity for an HA from a Group 1 influenzastrain, e.g., an H1, e.g., the H1 from an H1N1 strain, e.g., A/SouthCarolina/1/1918, A/Puerto Rico/08/1934, or A/California/04/2009, or anH5N1 strain, e.g., A/Indonesia/5/2005 or A/Vietnam/1203/2004, and/or anaffinity for an HA from a Group 2 influenza strain, e.g., an H3polypeptide, from an H3N2 strain, e.g., from A/Brisbane/59/2007); and b)a second K_(D) (an affinity for an influenza B HA), wherein the firstand second K_(D) are: one or both of: both equal to or less than 10⁻⁸nM; and within 10 or 100 fold of each other.

In an embodiment, the antibody molecule binds to at least one HApolypeptide from a Group 1 influenza strain with a higher affinity thana reference anti-HA antibody, and to at least one HA polypeptide from aGroup 2 influenza strain with a higher affinity than a reference anti-HAantibody. In another embodiment, the antibody molecule binds to at leastone HA polypeptide from an influenza A strain with a higher affinitythan a reference anti-HA antibody, and to at least one HA polypeptidefrom an influenza B strain with a higher affinity than a referenceanti-HA antibody. Exemplary reference HA antibodies include Ab 67-11(U.S. Provisional Application No. 61/645,453, U.S. ApplicationPublication No. 2013/0302348, and International Application PublicationNo. WO 2013/169377), FI6 (FI6, as used herein, refers to anyspecifically disclosed FI6 sequence in U.S. Published Application No.2010/0080813, U.S. published application No. 2011/0274702, WO2013/011347or Corti et al., Science 333:850-856, 2011, published online Jul. 28,2011; FIG. 4), FI28 (U.S. Published Application No. 2010/0080813), andC179 (Okuno et al., J. Virol. 67:2552-1558, 1993), F10 (Sui et al., Nat.Struct. Mol. Biol. 16:265, 2009), CR9114 (Dreyfus et al., Science. 2012;337(6100):1343-1348; published online Aug. 9, 2012), and CR6261 (Ekiertet al., Science 324:246-251, 2009).

Affinity, or relative affinity or aviditiy, can be measured by methodsknown in the art, such as by ELISA assay (Enzyme Linked ImmunosorbentAssay), Surface Plasmon Resonance (SPR, e.g., by a Biacore™ Assay), orKinExA® assay (Sapidyne, Inc.). Relative binding affinity is expressedherein according to ELISA assay. As used herein, an anti-HA antibodythat binds with “high affinity” to a Group 1 HA, to a Group 2 HA, and toa influenza B HA, can bind a Group 1 HA with a Kd less than or equal to200 pM, e.g., less than or equal to 100 pM, as measured by ELISA, canbind a Group 2 HA with a Kd less than or equal to 200 pM, e.g., lessthan or equal to 100 pM, as measured by ELISA, and can bind an influenzaB HA with a Kd less than or equal to 200 pM, e.g., less than or equal to100 pM, as measured by ELISA.

Exemplary Anti-HA Antibody Molecules

Provided herein are antibodies that have one or more CDR sequences andone or more framework (FR) sequences as shown in Table 2.

TABLE 2Heavy and Light Chain CDR and FR Sequences for Anti-HA Antibodies CDR/FRSEQ ID Region Amino Acid Sequence NO: HC CDR1 [S/T]Y[A/G]MH   1 HC CDR2V[I/V/L]S[Y/F]D/G[S/N][Y/N][K/R]YYADSVQG   2 HC CDR3D[S/T][R/K/Q]LR[S/T]LLYFEWLS[Q/S]G[Y/L/V][F/L][N/D][P/Y]   3 LC CDR1Q[S/T][V/L/I][T/S][Y/F/W][N/S/D]YKNYLA   4 LC CDR1Q[S/T][V/L/I][T/S][Y/F/W][N/S/D/Q/R/E]YKNYLA 170 LC CDR2W[A/G]S[T/A/Y/H/K/D][R/L]E[S/T]   5 LC CDR3 QQ[Y/H]YRTPP[T/S]   6 HC FR1[E/Q]VQLLE[S/T]GGGLVKPGQSLKLSCAASGFTF[S/T]   7 HC FR2 WVRQPPGKGLEWVA   8HC FR3 RFTISRDNSKNTLYLQMNSLRAEDTAVYYCAK   9 HC FR4WG[A/Q]G[T/A][T/M][L/V]TVSS  10 LC FR1[E/D]I[V/Q]MTQSP[D/S][S/T]L/V][A/S][V/A][S/T]L/V/R]G[E/D]R  11[A/V][T/S]I[N/T/Q/D/R/]C[K/R]SS LC FR2 WYQQKPG[Q/K][P/A]PKLLIY  12LC FR3 GVP[D/E/S]RFSGSGSGTDFTLTISSLQ[A/P]ED[V/F/K/D]A[V/T]YYC  13 LC FR4FG[G/Q/T/S/N]GTK[L/V]IK  14

In an embodiment, the anti-HA antibody comprises a heavy chain and/or alight chain as defined in Table 3 below. The amino acid sequences of thevariable heavy and light chains of Table 3 are provided in FIGS. 2, 3,respectively, or in FIG. 7.

TABLE 3 Heavy and Light Chain Amino Acid Sequence Designations forAnti-HA Antibodies Antibody HC SEQ ID NO: LC SEQ ID NO: 1. Ab A18 15 1528 28 2. Ab 014 16 16 29 29 3. Ab 028 16 16 30 30 4. Ab 001 17 17 31 315. Ab 002 18 18 31 31 6. Ab 003 19 19 31 31 7. Ab 009 17 17 32 32 8. Ab010 18 18 32 32 9. Ab 011 19 19 32 32 10. Ab 017 17 17 33 33 11. Ab B1818 18 33 33 12. Ab 019 19 19 33 33 13. Ab 025 17 17 34 34 14. Ab 026 1818 34 34 15. Ab 027 19 19 34 34 16. Ab 086 20 20 34 34 17. Ab 154 21 2129 29 18. Ab 155 21 21 30 30 19. Ab 157 22 22 29 29 20. Ab 159 22 22 3535 21. Ab 160 17 17 36 36 22. Ab 186 17 17 37 37 23. Ab 187 17 17 38 3824. Ab 188 17 17 39 39 25. Ab 189 17 17 40 40 26. Ab 190 17 17 41 41 27.Ab 191 17 17 42 42 28. Ab 192 17 17 43 43 29. Ab 193 17 17 44 44 30. Ab194 19 19 37 37 31. Ab 195 19 19 38 38 32. Ab 196 19 19 39 39 33. Ab 19719 19 40 40 34. Ab 198 19 19 41 41 35. Ab 199 19 19 42 42 36. Ab 200 1919 43 43 37. Ab 202 17 17 45 45 38. Ab 203 18 18 45 45 39. Ab 204 19 1945 45 40. Ab 210 23 23 45 45 41. Ab 211 17 17 46 46 42. Ab 212 18 18 4646 43. Ab 213 19 19 46 46 44. Ab 219 23 23 46 46 45. Ab A001 24 24 47 4746. Ab A002 24 24 48 48 47. Ab A003 24 24 49 49 48. Ab 004 25 25 47 4749. Ab 005 25 25 48 48 50. Ab 006 25 25 49 49 51. Ab 007 26 26 47 47 52.Ab 008 26 26 48 48 53. Ab A009 26 26 49 49 54. Ab A010 24 24 50 50 55.Ab A011 24 24 51 51 56. Ab 012 25 25 50 50 57. Ab 013 25 25 51 51 58. AbA14 26 26 50 50 59. Ab 015 26 26 51 51 60. Ab 016 27 27 47 47 61. AbA017 27 27 48 48 62. Ab C18 27 27 49 49 63. Ab A019 27 27 50 50 64. Ab031 24 24 45 45 65. Ab 032 25 25 45 45 66. Ab 033 26 26 45 45 67. Ab 03427 27 45 45 68. Ab 037 24 24 46 46 69. Ab 038 25 25 46 46 70. Ab 039 2626 46 46 71. Ab 040 27 27 46 46 72. Ab 043 25 25 60 60 73. Ab 044 25 2552 52 74. Ab 045 25 25 57 57 75. Ab 046 25 25 59 59 76. Ab 047 25 25 5555 77. Ab 048 25 25 58 58 78. Ab 049 25 25 54 54 79. Ab 050 25 25 56 5680. Ab 051 25 25 53 53 81. Ab 052 25 25 61 61 82. Ab 067 25 25 153 15383. Ab 068 25 25 154 154 84. Ab 069 25 25 155 155 85. Ab 070 25 25 156156 86. Ab 071 162 162 52 52 87. Ab 072 163 163 52 52 88. Ab 073 25 25165 165 89. Ab 074 25 25 166 166 90. Ab 075 25 25 167 167 91. Ab 076 2525 168 168 92. Ab 077 25 25 169 169 93. Ab 078 164 164 52 52 94. Ab 079164 164 155 155 95. Ab 080 164 164 166 166 96. Ab 081 164 164 169 169 AbA18 is also sometimes known as Ab 018 herein.

In an embodiment, the anti-HA antibody comprises a heavy chain asdefined in Table 4A below, and/or a light chain as defined in Table 4Abelow.

TABLE 4A Heavy and Light Chain Amino Acid Sequence Designations HC SEQID NO: LC SEQ ID NO: 15 15 28 28 16 16 29 29 17 17 30 30 18 18 35 35 1919 31 31 21 21 32 32 22 22 33 33 20 20 34 34 23 23 36 36 24 24 45 45 2525 46 46 26 26 37 37 27 27 38 38 Hc consensus 161 39 39 (HC161) 162 16240 40 163 163 41 41 164 164 42 42 43 43 44 44 47 47 48 48 49 49 50 50 5151 52 52 53 53 54 54 55 55 56 56 57 57 58 58 59 59 60 60 61 61 153 153154 154 155 155 156 156 LC consensus 62 (LC62) 165 165 166 166 167 167168 168 169 169

In an embodiment, an antibody molecule described herein comprises aheavy chain sequence as defined in Table 4A and a light chain sequenceas defined in Table 4A.

In an embodiment, an antibody molecule described herein comprises aheavy chain sequence as defined herein, e.g., in Table 4A, where adipeptide is fused to the N-terminus. Typically, the dipeptide isisoleucine-aspartic acid (Ile-Asp). In another embodiment, an antibodymolecule described herein comprises a light chain sequence as definedherein, e.g., in Table 4A, where a dipeptide is fused to the N-terminus.Typically, the dipeptide is Ile-Asp. In yet another embodiment, anantibody molecule described herein comprises a heavy chain comprising anN-terminal Ile-Asp dipeptide and a light chain comprising an Ile-Aspdipeptide. In the propeptide sequence of the heavy chain or light chainpolypeptide, the Ile-Asp dipeptide occurs between the signal sequenceand FR1. Heavy chain and light chain variable sequences comprising anIle-Asp dipeptide at the N-terminus are identified in Table 4B.

TABLE 4B Heavy and Light Chain Amino Acid Sequence Designations, wherethe Sequence Includes an N-terminal Ile-Asp Dipeptide HC SEQ ID NO: LCSEQ ID NO: 15-ID 96 28-ID 110 16-ID 97 29-ID 111 17-ID 98 30-ID 11218-ID 99 35-ID 113 19-ID 100 31-ID 114 21-ID 101 32-ID 115 22-ID 10233-ID 116 20-ID 103 34-ID 117 23-ID 104 36-ID 118 24-ID 105 45-ID 11925-ID 106 46-ID 120 26-ID 107 37-ID 121 27-ID 108 38-ID 122 Hc consensus109 39-ID 123 ID (161-ID) 40-ID 124 41-ID 125 42-ID 126 43-ID 127 44-ID128 47-ID 129 48-ID 130 49-ID 131 50-ID 132 51-ID 133 52-ID 134 53-ID135 54-ID 136 55-ID 137 56-ID 138 57-ID 139 58-ID 140 59-ID 141 60-ID142 61ID  143 153-ID  157 154-ID  158 155-ID  159 156-ID  160 LCconsensus 144 ID (62-ID)

In another embodiment, an antibody molecule described herein is otherthan an antibody known in the art. For example, the antibody is not Ab67-11 (U.S. Provisional Application No. 61/645,453, U.S. ApplicationPublication No. 2013/0302348, and International Application PublicationNo. WO 2013/169377), FI6 (FI6, as used herein, refers to anyspecifically disclosed FI6 sequence in U.S. Application Publication No.2010/0080813, U.S. Application Publication No. 2011/0274702,WO2013/011347 or Corti et al., Science 333:850-856, 2011, publishedonline Jul. 28, 2011; FIGS. 12A to 12C), FI28 (U.S. ApplicationPublication No. 2010/0080813), C179 (Okuno et al., J. Virol. 67:2552,1993), F10 (Sui et al., Nat. Struct. Mol. Biol. 16:265, 2009), CR9114(Dreyfus et al., Science 337:1343, 2012), or CR6261 (Ekiert et al.,Science 324:246, 2009). In an embodiment, an antibody described hereinis other than Ab 67-11 (U.S. Provisional Application No. 61/645,453,U.S. Application Publication No. 2013/0302348, and InternationalApplication Publication No. WO 2013/169377).

Variants

In an embodiment, an antibody molecule described herein has a variableheavy chain immunoglobulin domain that is at least 85%, 87%, 88%, 89%,90%, 92%, 94%, 95%, 96%, 97%, 98%, or 99% homologous, or at least 85%,87%, 88%, 89%, 90%, 92%, 94%, 95%, 96%, 97%, 98%, or 99% identical, to aheavy chain disclosed herein, e.g., from Table 3, Table 4A, Table 4B,FIG. 2, FIG. 5 or FIG. 7, e.g. consensus sequence of SEQ ID NO:161, andhas a variable light chain immunoglobulin domain that is at least 85%,87%, 88%, 89%, 90%, 92%, 94%, 95%, 96%, 97%, 98%, or 99% homologous, orat least 85%, 87%, 88%, 89%, 90%, 92%, 94%, 95%, 96%, 97%, 98%, or 99%identical, to a light chain disclosed herein, e.g., from Table 3, Table4A, Table 4B, FIGS. 3A-3B, FIGS. 6A-6B or FIG. 7, e.g., the consensussequence of SEQ ID NO:62. The consensus sequences were determinedthrough the analysis of biochemical and biophysical properties ofseveral hundred computationally designed VH/VL combinations. Theconsensus sequences represent the amino acid sequences in which eachamino acid is the one that occurs most frequently at that site whenmultiple sequences comprising desirable biochemical and biophysical dataare aligned.

An exemplary anti-HA binding antibody has one or more CDRs, e.g., allthree HC CDRs and/or all three LC CDRs of a particular antibodydisclosed herein, or CDRs that are, in sum, at least 85%, 87%, 88%, 89%,90%, 92%, 94%, 95%, 96%, 97%, 98%, or 99% homologous, or at least 85%,87%, 88%, 89%, 90%, 92%, 94%, 95%, 96%, 97%, 98%, or 99% identical, tosuch an antibody.

In an embodiment, the H1 and H2 hypervariable loops have the samecanonical structure as those of an antibody described herein. In anembodiment, the L1 and L2 hypervariable loops have the same canonicalstructure as those of an antibody described herein.

In an embodiment, the amino acid sequence of the HC and/or LC variabledomain sequence is at least 85%, 87%, 88%, 89%, 90%, 92%, 94%, 95%, 96%,97%, 98%, or 99% homologous, or at least 85%, 87%, 88%, 89%, 90%, 92%,94%, 95%, 96%, 97%, 98%, or 99% identical, to the amino acid sequence ofthe HC and/or LC variable domain of an antibody described herein. Theamino acid sequence of the HC and/or LC variable domain sequence candiffer by at least one amino acid, but no more than ten, eight, six,five, four, three, or two amino acids from the corresponding sequence ofan antibody described herein. For example, the differences may beprimarily or entirely in the framework regions.

In certain embodiments, the amino acid differences are conservativeamino acid differences (e.g., conservative amino acid substitutions). A“conservative” amino acid substitution is one in which the amino acidresidue is replaced with an amino acid residue comprising a similar sidechain. Families of amino acid residues comprising similar side chainshave been defined in the art. These families include, e.g., amino acidswith basic side chains (e.g., lysine, arginine, histidine), acidic sidechains (e.g., aspartic acid, glutamic acid), uncharged polar side chains(e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine,cysteine), nonpolar side chains (e.g., alanine, valine, leucine,isoleucine, proline, phenylalanine, methionine, tryptophan),beta-branched side chains (e.g., threonine, valine, isoleucine) andaromatic side chains (e.g., tyrosine, phenylalanine, tryptophan,histidine).

The amino acid sequences of the HC and LC variable domain sequences canbe encoded by a nucleic acid sequence that hybridizes under highstringency conditions to a nucleic acid sequence described herein or onethat encodes a variable domain or an amino acid sequence describedherein. In an embodiment, the amino acid sequences of one or moreframework regions (e.g., FR1, FR2, FR3, and/or FR4) of the HC and/or LCvariable domain are at least 85%, 87%, 88%, 89%, 90%, 92%, 94%, 95%,96%, 97%, 98%, or 99% homologous, or at least 85%, 87%, 88%, 89%, 90%,92%, 94%, 95%, 96%, 97%, 98%, or 99% identical, to correspondingframework regions of the HC and LC variable domains of an antibodydescribed herein. In an embodiment, one or more heavy or light chainframework regions (e.g., HC FR1, FR2, and FR3) are at least 85%, 87%,88%, 89%, 90%, 92%, 94%, 95%, 96%, 97%, 98%, or 99% homologous, or atleast 85%, 87%, 88%, 89%, 90%, 92%, 94%, 95%, 96%, 97%, 98%, or 99%identical, to the sequence of corresponding framework regions from ahuman germline antibody.

Production of Binding Agents

Nucleic acids (e.g., the genes) encoding a binding agent, e.g., anantibody molecule, generated by a method described herein can besequenced, and all or part of the nucleic acids can be cloned into avector that expresses all or part of the nucleic acids. For example, thenucleic acids can include a fragment of the gene encoding the antibody,such as a single chain antibody (scFv), a F(ab′)₂ fragment, a Fabfragment, or an Fd fragment.

The disclosure also provides host cells comprising the nucleic acidsencoding an antibody or fragment thereof as described herein. The hostcells can be, for example, prokaryotic or eukaryotic cells, e.g.,mammalian cells, or yeast cells, e.g., Pichia (see e.g., Powers et al.(2001) J. Immunol. Methods 251:123-35), Hanseula, or Saccharomyces.

Antibody molecules, particularly full length antibody molecules, e.g.,IgGs, can be produced in mammalian cells. Exemplary mammalian host cellsfor recombinant expression include Chinese Hamster Ovary (CHO) cells(including dhfr⁻ CHO cells, described in Urlaub and Chasin (1980) Proc.Natl. Acad. Sci. USA 77:4216-4220, used with a DHFR selectable marker,e.g., as described in Kaufman and Sharp (1982) Mol. Biol. 159:601-621),lymphocytic cell lines, e.g., NSO myeloma cells and SP2 cells, COScells, K562, and a cell from a transgenic animal, e.g., a transgenicmammal. For example, the cell is a mammary epithelial cell.

In addition to the nucleic acid sequence encoding the immunoglobulindomain, the recombinant expression vectors may carry additional nucleicacid sequences, such as sequences that regulate replication of thevector in host cells (e.g., origins of replication) and selectablemarker genes. The selectable marker gene facilitates selection of hostcells into which the vector has been introduced (see e.g., U.S. Pat.Nos. 4,399,216; 4,634,665; and 5,179,017). Exemplary selectable markergenes include the dihydrofolate reductase (DHFR) gene (for use in dhfr⁻host cells with methotrexate selection/amplification) and the neo gene(for G418 selection).

In an exemplary system for recombinant expression of an antibodymolecule (e.g., a full length antibody or an antigen-binding portionthereof), a recombinant expression vector encoding both the antibodyheavy chain and the antibody light chain is introduced into dhfr-CHOcells by calcium phosphate-mediated transfection. Within the recombinantexpression vector, the antibody heavy and light chain genes are eachoperatively linked to enhancer/promoter regulatory elements (e.g.,derived from SV40, CMV, adenovirus and the like, such as a CMVenhancer/AdMLP promoter regulatory element or an SV40 enhancer/AdMLPpromoter regulatory element) to drive high levels of transcription ofthe genes. The recombinant expression vector also carries a DHFR gene,which allows for selection of CHO cells that have been transfected withthe vector using methotrexate selection/amplification. The selectedtransformant host cells are cultured to allow for expression of theantibody heavy and light chains and intact antibody molecule isrecovered from the culture medium. Standard molecular biology techniquesare used to prepare the recombinant expression vector, to transfect thehost cells, to select for transformants, to culture the host cells, andto recover the antibody from the culture medium. For example, someantibodies can be isolated by affinity chromatography with a Protein Aor Protein G. For example, purified antibodies can be concentrated toabout 100 mg/mL to about 200 mg/mL using protein concentrationtechniques that are known in the art.

Antibody molecules can also be produced by a transgenic animal. Forexample, U.S. Pat. No. 5,849,992 describes a method for expressing anantibody molecule in the mammary gland of a transgenic mammal. Atransgene is constructed that includes a milk-specific promoter andnucleic acid sequences encoding the antibody molecule of interest, e.g.,an antibody described herein, and a signal sequence for secretion. Themilk produced by females of such transgenic mammals includes, secretedtherein, the antibody of interest, e.g., an antibody described herein.The antibody molecule can be purified from the milk, or for someapplications, used directly.

Antibody molecules can also be expressed in vivo, followingadministration of a vector containing nucleic acids encoding theantibody heavy chain and the antibody light chain. Vector mediatedgene-transfer is then used to engineer secretion of the anti-HA antibodyinto circulation. For example, an anti-HA antibody heavy chain and ananti-HA antibody light chain as described herein are cloned into anadeno-associated virus (AAV)-based vector, and each of the anti-HAantibody heavy chain and the anti-HA antibody light chain are undercontrol of a promoter, such as a cytomegalovirus (CMV) promoter.Administration of the vector to a subject, such as to a patient, e.g., ahuman patient, such as by intramuscular injection, results in expressionof an anti-HA antibody, and secretion into the circulation.

Modifications of Binding Agents

Binding, agents, e.g., antibody molecules, described herein, can bemodified to have numerous properties, e.g., to have altered, e.g.,extended half life, to be associated with, e.g., covalently bound todetectable moieties, e.g., labels, to be associated with, e.g.,covalently bound to toxins, or to have other properties, e.g., alteredimmune fucntions.

Antibody molecules may include modifications, e.g., modifications thatalter Fc function, e.g., to decrease or remove interaction with an Fcreceptor or with C1q, or both. In one example, the human IgG1 constantregion can be mutated at one or more residues.

For some antibody molecules that include an Fc domain, the antibodyproduction system may be designed to synthesize antibody molecules inwhich the Fc region is glycosylated. The Fc domain can be produced in amammalian expression system that appropriately glycosylates the residuecorresponding to asparagine 297. The Fc domain can also include othereukaryotic post-translational modifications.

Other suitable Fc domain modifications include those described inWO2004/029207. For example, the Fc domain can be an XmAb® Fc (Xencor,Monrovia, Calif.). The Fc domain, or a fragment thereof, can have asubstitution in an Fcγ Receptor (FcγR) binding region, such as thedomains and fragments described in WO05/063815. In some embodiments, theFc domain, or a fragment thereof, has a substitution in a neonatal FcReceptor (FcRn) binding region, such as the domains and fragmentsdescribed in WO05047327. In other embodiments, the Fc domain is a singlechain, or fragment thereof, or modified version thereof, such as thosedescribed in WO2008143954. Other suitable Fc modifications are known anddescribed in the art.

Antibody molecules can be modified, e.g., with a moiety that improvesits stabilization and/or retention in circulation, e.g., in blood,serum, lymph, bronchoalveolar lavage, or other tissues, e.g., by atleast 1.5, 2, 5, 10, or 50 fold.

For example, an antibody molecule generated by a method described hereincan be associated with a polymer, e.g., a substantially non-antigenicpolymer, such as a polyalkylene oxide or a polyethylene oxide. Suitablepolymers will vary substantially by weight. Polymers comprisingmolecular number average weights ranging from about 200 to about 35,000daltons (or about 1,000 to about 15,000, and 2,000 to about 12,500) canbe used.

For example, an antibody molecule generated by a method described hereincan be conjugated to a water soluble polymer, e.g., a hydrophilicpolyvinyl polymer, e.g. polyvinylalcohol or polyvinylpyrrolidone. Anon-limiting list of such polymers include polyalkylene oxidehomopolymers such as polyethylene glycol (PEG) or polypropylene glycols,polyoxyethylenated polyols, copolymers thereof and block copolymersthereof, provided that the water solubility of the block copolymers ismaintained. Additional useful polymers include polyoxyalkylenes such aspolyoxyethylene, polyoxypropylene, and block copolymers ofpolyoxyethylene and polyoxypropylene (Pluronics); polymethacrylates;carbomers; branched or unbranched polysaccharides that comprise thesaccharide monomers D-mannose, D- and L-galactose, fucose, fructose,D-xylose, L-arabinose, D-glucuronic acid, sialic acid, D-galacturonicacid, D-mannuronic acid (e.g. polymannuronic acid, or alginic acid),D-glucosamine, D-galactosamine, D-glucose and neuraminic acid includinghomopolysaccharides and heteropolysaccharides such as lactose,amylopectin, starch, hydroxyethyl starch, amylose, dextrane sulfate,dextran, dextrins, glycogen, or the polysaccharide subunit of acidmucopolysaccharides, e.g. hyaluronic acid; polymers of sugar alcoholssuch as polysorbitol and polymannitol; heparin or heparan.

Binding agents, e.g., antibody molecules, as disclosed herein, can byconjugated to another entity or moiety (e.g., to a cytotoxic orcytostatic moiety, a label or detectable moiety, or a therapeuticmoiety). Exemplary moieties include: a cytotoxic or cytostatic agent,e.g., a therapeutic agent, a drug, a compound emitting radiation,molecules of plant, fungal, or bacterial origin, or a biological protein(e.g., a protein toxin) or particle (e.g., a recombinant viral particle,e.g., via a viral coat protein), a detectable agent; a pharmaceuticalagent, and/or a protein or peptide that can mediate association of theantibody or antibody portion with another molecule (such as astreptavidin core region or a polyhistidine tag). A binding agent, e.g.,an antibody molecule, as disclosed herein, can be functionally linked byany suitable method (e.g., chemical coupling, genetic fusion, covalentbinding, noncovalent association or otherwise) to one or more othermolecular entities.

Binding agents, e.g., antibody molecules, disclosed herein can beconjugated with a detectable moiety, e.g., a label or imaging agent.Such moieties can include enzymes (e.g., horseradish peroxidase,beta-galactosidase, luciferase, alkaline phosphatase,acetylcholinesterase, glucose oxidase and the like), radiolabels (e.g.,³H, ¹⁴C, ¹⁵N, ³⁵S, ⁹⁰Y, ⁹⁹Tc, ¹¹¹In, ¹²⁵I, ¹³¹I and the like), haptens,fluorescent labels (e.g., FITC, rhodamine, lanthanide phosphors,fluorescein, fluorescein isothiocyanate, rhodamine,5-dimethylamine-1-napthalenesulfonyl chloride, phycoerythrin and thelike), phosphorescent molecules, chemiluminescent molecules,chromophores, luminescent molecules, photoaffinity molecules, coloredparticles or affinity ligands, such as biotin, predetermined polypeptideepitopes recognized by a secondary reporter (e.g., leucine zipper pairsequences, or binding sites for secondary antibodies, metal bindingdomains, epitope tags). In some embodiments, a moiety, e.g., adetectable moiety, e.g., a label, is attached by spacer arms of variouslengths to reduce potential steric hindrance.

In an embodiment, a binding agent, e.g., antibody molecule, disclosedherein, is derivatized with a detectable enzyme and is detected byadding additional reagents that the enzyme uses to produce a detectablereaction product. For example, when the detectable agent horseradishperoxidase is present, the addition of hydrogen peroxide anddiaminobenzidine leads to a colored reaction product, which isdetectable. A binding agent, e.g., antibody molecule, disclosed herein,ay also be derivatized with a prosthetic group (e.g.,streptavidin/biotin and avidin/biotin). For example, an antibody may bederivatized with biotin, and detected through indirect measurement ofavidin or streptavidin binding.

In an embodiment, the moiety comprises paramagnetic ions andNMR-detectable substances, among others. For example, in someembodiments, a paramagnetic ion is one or more of chromium (III),manganese (II), iron (III), iron (II), cobalt (II), nickel (II), copper(II), neodymium (III), samarium (III), ytterbium (III), gadolinium(III), vanadium (II), terbium (III), dysprosium (III), holmium (III),erbium (III), lanthanum (III), gold (III), lead (II), and/or bismuth(III).

Binding agents, e.g., antibody molecules, as disclosed herein, can bemodified to be associated with, e.g., conjugated to, a therapeuticagent, e.g., an agent comprising anti-viral activity, anti-inflammatoryactivity, or cytotoxic activity, etc. In some embodiments, therapeuticagents can treat symptoms or causes of influenza infection (e.g., forexample, anti-viral, pain-relief, anti-inflammatory, immunomodulatory,sleep-inducing activities, etc.).

Treatment Methods and Administration

The binding agents, e.g., antibody molecules, or formulations thereof,featured in the disclosure, can be used to treat a subject, e.g., asubject, e.g., a human subject, infected with, or at risk for becominginfected with, an influenza virus.

Any human is candidate to receive an antibody molecule disclosed hereinfor treatment or prevention of an infection by an influenza virus.Humans at high risk of infection, such as immunocompromised individuals,and humans who are at high risk of exposure to influenza virus areparticularly suited to receive treatment with the antibody molecule.Immunocompromised individuals include the elderly (65 years and older)and children (e.g., 6 months to 18 years old), and people with chronicmedical conditions. People at high risk of exposure include heath careworkers, teachers and emergency responders (e.g., firefighters,policemen).

The antibody molecules described herein can also be used to prevent orreduce (e.g., minimize) secondary infection (e.g., secondary bacterialinfection) or a risk of comprising secondary infection associated withinfluenza, or any effects (e.g., symptoms or complications) thereof on asubject. Opportunistic secondary bacterial infections (e.g., secondarybacterial pneumonia, e.g., primarily with Streptococcus pneumonia)contribute significantly to the overall morbidity and mortalityassociated with seasonal and pandemic influenza infections. The antibodymolecules described herein can be used to prevent or reduce (e.g.,minimize) the complications from secondary, opportunistic infections(e.g., bacterial infections) in a subject.

An antibody molecule can be administered to a subject, e.g., a humansubject, by a variety of methods. For many applications, the route ofadministration is one of: intravenous injection or infusion,subcutaneous injection, or intramuscular injection. An antibody moleculecan be administered as a fixed dose, or in a mg/kg dose. The antibodymolecule can be administered intravenously (IV) or subcutaneously (SC).For example, the antibody molecule can be administered at a fixed unitdose of between about 50-600 mg IV, e.g., every 4 weeks, or betweenabout 50-100 mg SC (e.g., 75 mg), e.g., at least once a week (e.g.,twice a week). In an embodiment, the antibody molecule is administeredIV at a fixed unit dose of 50 mg to 10000 mg, e.g., 1000 mg to 5000 mg,2000 mg to 5000 mg, 2000 mg to 3000 mg, 2300 to 4600 mg, or 4000 mg to5000 mg, e.g., 50 mg, 60 mg, 80 mg, 100 mg, 120 mg, 130 mg, 140 mg, 150mg, 160 mg, 180 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 800mg, 900 mg, 1000 mg, 1100 mg, 1200 mg, 1300 mg, 1400 mg, 1500 mg, 1600mg, 1700 mg, 1800 mg, 1900 mg, 2000 mg, 2100 mg, 2200 mg, 2300 mg, 2400mg, 2500 mg, 2600 mg, 2700 mg, 2800 mg, 2900 mg, 3000 mg, 3100 mg, 3200mg, 3300 mg, 3400 mg, 3500 mg, 3600 mg, 3700 mg, 3800 mg, 3900 mg, 4000mg, 4100 mg, 4200 mg, 4300 mg, 4400 mg, 4500 mg, or more. Administrationof the IV dose can be once or twice or three times or more per week, oronce every two, three, four, or five weeks, or less frequently.

In an embodiment, the antibody molecule is administered SC at a fixedunit dose of 50 mg, 60 mg, 70 mg, 75 mg, 80 mg, 100 mg, or 120 mg ormore. Administration of the SC dose can be once or twice or three timesor more per week, or once every two, three, four, or five weeks, or lessfrequently.

An anti-HA antibody molecule disclosed herein can also be administeredby inhalation, such as by intranasal or by oral inhalation, such as at afixed unit dose of 50 mg, 60 mg, 80 mg, 100 mg, 120 mg, 130 mg, 140 mg,150 mg, 160 mg, 180 mg, 200 mg, 300 mg, 400 mg, 500 mg, or 600 mg ormore.

In an embodiment, an anti-HA antibody is administered to a subject viavector-mediated gene transfer, such as through the delivery of a vectorencoding the heavy chain and the light chain of an anti-HA antibody, andthe antibody is expressed from the heavy chain and light chain genes inthe body. For example, nucleic acids encoding a heavy chain and a lightchain can be cloned in a AAV vector, such as a self-complementary AAVvector, the scAAV vector administered to a human by injection, such asby IM injection, and the antibody is expressed and secreted into thecirculation of the human.

An antibody molecule can also be administered in a bolus at a dose ofbetween about 1 and 50 mg/kg, e.g., between about 1 and 10 mg/kg,between about 1 and 25 mg/kg or about 25 and 50 mg/kg, e.g., about 50mg/kg, 25 mg/kg, 10 mg/kg, 6.0 mg/kg, 5.0 mg/kg, 4.0 mg/kg, 3.0 mg/kg,2.0 mg/kg, 1.0 mg/kg, or less. Modified dose ranges include a dose thatis less than about 3000 mg/subject, about 1500 mg/subject, about 1000mg/subject, about 600 mg/subject, about 500 mg/subject, about 400mg/subject, about 300 mg/subject, about 250 mg/subject, about 200mg/subject, or about 150 mg/subject, typically for administration everyfourth week or once a month. The antibody molecule can be administered,for example, every three to five weeks, e.g., every fourth week, ormonthly.

Dosing can be adjusted according to a patient's rate of clearance of aprior administration of the antibody. For example, a patient may not beadministered a second or follow-on dose before the level of antibodiesin the patient's system has dropped below a pre-determined level. In anembodiment, a sample from a patient (e.g., plasma, serum, blood, urine,or cerebrospinal fluid (CSF)) is assayed for the presence of antibodies,and if the level of antibodies is above a pre-determined level, thepatient will not be administered a second or follow-on dose. If thelevel of antibodies in the patient's system is below a pre-determinedlevel, then the patient is administered a second or follow-on dose. Apatient whose antibody levels are determined to be too high (above thepre-determined level) can be tested again after one or two or threedays, or a week, and if the level of antibody in the patient samples hasdropped below the pre-determined level, the patient may be administereda second or follow-on dose of antibody.

In certain embodiments, the antibody may be prepared with a carrier thatwill protect the drug against rapid release, such as a controlledrelease formulation, including implants, and microencapsulated deliverysystems. Biodegradable, biocompatible polymers can be used, such asethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen,polyorthoesters, and polylactic acid. Many methods for the preparationof such formulations are patented or generally known. See, e.g.,Controlled Drug Delivery (Drugs and the Pharmaceutical Sciences), SecondEdition, J. Robinson and V. H. L. Lee, eds., Marcel Dekker, Inc., NewYork, 1987.

Pharmaceutical compositions can be administered with a medical device.For example, pharmaceutical compositions can be administered with aneedleless hypodermic injection device, such as the devices disclosed inU.S. Pat. Nos. 5,399,163; 5,383,851; 5,312,335; 5,064,413; 4,941,880;4,790,824; or 4,596,556. Examples of well-known implants and modules arediscussed in, e.g., U.S. Pat. No. 4,487,603, which discloses animplantable micro-infusion pump for dispensing medication at acontrolled rate; U.S. Pat. No. 4,486,194, which discloses a therapeuticdevice for administering medicaments through the skin; U.S. Pat. No.4,447,233, which discloses a medication infusion pump for deliveringmedication at a precise infusion rate; U.S. Pat. No. 4,447,224, whichdiscloses a variable flow implantable infusion apparatus for continuousdrug delivery; U.S. Pat. No. 4,439,196, which discloses an osmotic drugdelivery system comprising multi-chamber compartments; and U.S. Pat. No.4,475,196, which discloses an osmotic drug delivery system. Of course,many other such implants, delivery systems, and modules are also known.

In an embodiment, the binding agent, e.g., an antibody molecule, isadministered buccally, orally, or by nasal delivery, e.g., as a liquid,spray, or aerosol, e.g., by topical application, e.g., by a liquid ordrops, or by inhalation.

An antibody molecule described herein can be administered with one ormore additional therapeutic agents, e.g., a second drug, for treatmentof a viral infection, or a symptom of the infection. The antibodymolecule and the one or more second or additional agents can beformulated together, in the same formulation, or they can be in separateformulations, and administered to a patient simultaneously orsequentially, in either order.

Dosage regimens are adjusted to provide the desired response, such as atherapeutic response or a combinatorial therapeutic effect. Generally,any combination of doses (either separate or co-formulated) of anantibody molecule and a second or additional agent can be used in orderto provide a subject with both agents in bioavailable quantities.

Dosage unit form or “fixed dose” as used herein refers to physicallydiscrete units suited as unitary dosages for the subjects to be treated;each unit contains a predetermined quantity of active compoundcalculated to produce the desired therapeutic effect in association withthe required pharmaceutical carrier and optionally in association withanother agent.

A pharmaceutical composition may include a “therapeutically effectiveamount” of an agent described herein. In an embodiment, where theantibody molecule is administered in combination with a second oradditional agent, such effective amounts can be determined based on thecombinatorial effect of the administered first and second or additionalagent. A therapeutically effective amount of an agent may also varyaccording to factors such as the disease state, age, sex, and weight ofthe individual, and the ability of the compound to elicit a desiredresponse in the individual, such as amelioration of at least oneinfection parameter, or amelioration of at least one symptom of theinfection, such as chills, fever, sore throat, muscle pain, headache,coughing, weakness, fatigue and general discomfort. A therapeuticallyeffective amount is also one in which any toxic or detrimental effectsof the composition are outweighed by the therapeutically beneficialeffects.

In an embodiment, administration of a binding agent, e.g., antibodymolecule, provided, e.g., as a pharmaceutical preparation, is by one ofthe following routes: oral, intravenous, intramuscular, intra-arterial,subcutaneous, intraventricular, transdermal, interdermal, rectal,intravaginal, intraperitoneal, topical (as by liquids, powders,ointments, creams, sprays, or drops), mucosal, nasal, buccal, enteral,sublingual; intratracheal instillation, bronchial instillation, and/orinhalation; and/or as an oral spray, nasal spray, and/or aerosol.

Combination Treatments and Exemplary Second or Additional Agents

Binding agents, e.g., antibody molecules, provided e.g., as formulations(e.g., pharmaceutical formulations), can be administered either alone orin combination with one or more other therapy, e.g., the administrationof a second or additional therapeutic agent.

In an embodiment, the combination can result in a lower dose of theantibody molecule or of the other therapy being needed, which, in anembodiment can reduce side effects. In an embodiment, the combinationcan result in enhanced delivery or efficacy of one or both agents. Theagents or therapies can be administered at the same time (e.g., as asingle formulation that is administered to a patient or as two separateformulations administered concurrently) or sequentially in any order.

Such second or additional agents include vaccines, anti-viral agents,and/or additional antibodies. In typical embodiments the second oradditional agent is not co-formulated with the binding agent, e.g.,antibody molecule, though in others it is.

In an embodiment, the binding agent, e.g., antibody molecule, and thesecond or additional agent are administered such that one or more of thefollowing is achieved: therapeutic levels, or therapeutic effects, ofone overlap the other; detectable levels of both are present at the sametime; or the therapeutic effect is greater than what would be seen inthe absence of either the binding agent, e.g., antibody molecule, or thesecond or additional agent. In an embodiment, each agent will beadministered at a dose and on a time schedule determined for that agent.

The second or additional agent can be, for example, for treatment orprevention of influenza. For example, the binding agents, e.g., antibodymolecules, e.g., therapeutic antibodies, provided herein can beadministered in combination with a vaccine, e.g., a vaccine describedherein or a mixture (a.k.a. a cocktail) of influenza peptides tostimulate the patient's immune system to prevent infection withparticular strains of influenza A. In other examples, the second oradditional agent is an anti-viral agent (e.g., an anti-NA or anti-M2agent), a pain reliever, an anti-inflammatory, an antibiotic, asteroidal agent, a second therapeutic antibody molecule (e.g., ananti-HA antibody), an adjuvant, a protease or glycosidase (e.g.,sialidase), etc.

Exemplary anti-viral agents include, e.g., vaccines, neuraminidaseinhibitors or nucleoside analogs. Exemplary anti-viral agents caninclude, e.g., zidovudine, gangcyclovir, vidarabine, idoxuridine,trifluridine, foscarnet, acyclovir, ribavirin, amantadine, remantidine,saquinavir, indinavir, ritonavir, alpha-interferons and otherinterferons, a neuraminidase inhibitor (e.g., zanamivir (Relenza®),oseltamivir (Tamiflu®), laninamivir, peramivir), rimantadine. Exemplarysecond antibody molecules include, for example Ab 67-11 (U.S.Provisional Application No. 61/645,453, U.S. Application Publication No.2013/0302348, and International Application Publication No. WO2013/169377), FI6 (U.S. Application Publication No. 2010/0080813), FI28(U.S. Application Publication No. 2010/0080813), C179 (Okuno et al., J.Virol. 67:2552-8, 1993), F10 (Sui et al., Nat. Struct. Mol. Biol.16:265, 2009), CR9114 (Dreyfus et al., Science 337:1343, 2012), orCR6261 (see e.g., Ekiert et al., Science 324:246, 2009). Thus, Ab 044can be used in combination of any of those antibodies. In otherembodiments, two or more binding agents, e.g., antibody moleculesdisclosed herein, can be administered in combination, e.g., Ab 044 canbe administered in combination with Ab 032. In the case of combinations,two agents can be administered as part of the same dosage unit oradministered separately. Other exemplary agents useful for treating thesymptoms associated with influenza infection are acetaminophen,ibuprofen, aspirin, and naproxen.

In an embodiment, the antibody molecule and the second or additionalagent are provided as a co-formulation, and the co-formulation isadministered to the subject. It is further possible, e.g., at least 24hours before or after administering the co-formulation, to administerseparately one dose of the antibody formulation and then one dose of aformulation containing a second or additional agent. In anotherimplementation, the antibody molecule and the second or additional agentare provided as separate formulations, and the step of administeringincludes sequentially administering the antibody molecule and the secondor additional agent. The sequential administrations can be provided onthe same day (e.g., within one hour of one another or at least 3, 6, or12 hours apart) or on different days.

In an embodiment, the antibody molecule and the second or additionalagent are each administered as a plurality of doses separated in time.The antibody molecule and the second or additional agent are generallyeach administered according to a regimen. The regimen for one or bothmay have a regular periodicity. The regimen for the antibody moleculecan have a different periodicity from the regimen for the second oradditional agent, e.g., one can be administered more frequently than theother. In one implementation, one of the antibody molecule and thesecond or additional agent is administered once weekly and the otheronce monthly. In another implementation, one of the antibody moleculeand the second or additional agent is administered continuously, e.g.,over a period of more than 30 minutes but less than 1, 2, 4, or 12hours, and the other is administered as a bolus. In an embodiment,sequential administrations are administered. The time betweenadministration of the one agent and another agent can be minutes, hours,days, or weeks. The use of an antibody molecule described herein canalso be used to reduce the dosage of another therapy, e.g., to reducethe side-effects associated with another agent that is beingadministered. Accordingly, a combination can include administering asecond or additional agent at a dosage at least 10, 20, 30, or 50% lowerthan would be used in the absence of the antibody molecule. The antibodymolecule and the second or additional agent can be administered by anyappropriate method, e.g., subcutaneously, intramuscularly, orintravenously.

In some embodiments, each of the antibody molecule and the second oradditional agent is administered at the same dose as each is prescribedfor monotherapy. In other embodiments, the antibody molecule isadministered at a dosage that is equal to or less than an amountrequired for efficacy if administered alone. Likewise, the second oradditional agent can be administered at a dosage that is equal to orless than an amount required for efficacy if administered alone.

In some cases, the formulations described herein, e.g., formulationscontaining an antibody molecule described herein, include one or moresecond or additional agents, or are administered in combination with aformulation containing one or more second or additional agents.

In an embodiment, a binding agent, e.g., antibody molecule, provided,e.g., as a pharmaceutical preparation, is administered by inhalation oraerosol delivery of a plurality of particles, e.g., particles comprisinga mean particle size of 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 microns.

In an embodiment, the formulation is used (e.g., administered) incombination with an immunogen or a vaccine. Exemplary immunogens andvaccines are described in International Application Publication No. WO2013/170139, the content of which is incorporated by reference in itsentirety.

Kits

A formulation (e.g., pharmaceutical formulation) disclosed herein, e.g.,generated by the methods described herein, can be provided (e.g.,packaged) in a kit. The kit can include one or more other components,e.g., containers, buffers or other diluents, delivery devices, and thelike.

In an embodiment, the kit includes materials for administering aformulation (e.g., pharmaceutical formulation) to a subject, such as fortreatment or prevention of infection by influenza viruses. For example,the kit can include one or more or all of: (a) a container that containsa formulation (e.g., pharmaceutical formulation) that includes anantibody molecule, optionally (b) a container that contains a secondtherapeutic agent, and optionally (c) informational material.

In another embodiment, the kit includes materials for using an antibodymolecule in a diagnostic assay, such as for detection of HA in abiological sample. For example, the kit can include one or more or allof: (a) a container that contains a formulation (e.g., pharmaceuticalformulation) that includes an antibody molecule, optionally (b) acontainer that contains a reagents, e.g., labeled with a detectablemoiety, to detect the antibody, e.g., for use in an ELISA orimmunohistochemistry assay, and optionally (c) informational material.In another embodiment, the kit comprises a formulation, e.g., a bindingagent (e.g., antibody molecule) comprising a detectable moiety.

In an embodiment, the kit comprises a solid substrate, e.g., bead,dipstick, array, and the like, on which is disposed a formulation, e.g.,a binding agent (e.g., antibody molecule).

The informational material can be descriptive, instructional, marketingor other material that relates to the methods described herein and/orthe use of the agents for therapeutic benefit, or for a diagnosticassay.

The informational material of the kits is not limited in its form. In anembodiment, the informational material can include information aboutproduction of the antibody, concentration, date of expiration, batch orproduction site information, and so forth. In an embodiment, theinformational material relates to methods of administering theformulation or antibody molecule, e.g., in a suitable dose, dosage form,or mode of administration (e.g., a dose, dosage form, or mode ofadministration described herein), to treat a subject who has aninfection, e.g., viral infection or secondary infection (e.g., secondarybacterial infection).

In another embodiment, the informational material relates to methods forusing the formulation or antibody molecule for a diagnostic assay, e.g.,to detect the presence of influenza viruses in a biological sample.

The information can be provided in a variety of formats, includingprinted text, computer readable material, video recording, or audiorecording, or information that provides a link or address to substantivematerial.

In addition to the binding agent (e.g., antibody molecule), theformulation in the kit can include other ingredients, such as a solventor buffer, a stabilizer, or a preservative. The binding agent (e.g.,antibody molecule) can be provided in any form, e.g., a liquid, dried orlyophilized form, and substantially pure and/or sterile. When the agentsare provided in a liquid solution, the liquid solution typically is anaqueous solution. When the agents are provided as a dried form,reconstitution generally is by the addition of a suitable solvent. Thesolvent, e.g., sterile water or buffer, can optionally be provided inthe kit.

The kit can include one or more containers for the formulationcontaining the binding agent. In an embodiment, the kit containsseparate containers, dividers or compartments for the formulation andinformational material. For example, the formulation can be contained ina bottle, vial, or syringe, and the informational material can becontained in a plastic sleeve or packet. In another embodiment, theseparate elements of the kit are contained within a single, undividedcontainer. For example, the formulation is contained in a bottle, vialor syringe that has attached thereto the informational material in theform of a label. In an embodiment, the kit includes a plurality (e.g., apack) of individual containers, each containing one or more unit dosageforms (e.g., a dosage form described herein) of the binding agent (e.g.,antibody molecule). The containers can include a combination unitdosage, e.g., a unit that includes both the antibody molecule and thesecond or additional agent, such as in a desired ratio. For example, thekit can include a plurality of syringes, ampoules, foil packets, blisterpacks, or medical devices each containing, for example, a singlecombination unit dose. The containers of the kits can be air tight,waterproof (e.g., impermeable to changes in moisture or evaporation),and/or light-tight.

In an embodiment, the kit comprises two containers, one of whichcontains the formulation (e.g., pharmaceutical formulation) and theother of which contains an adjuvant. In an embodiment, the kit comprisestwo containers, one of which contains the formulation (e.g.,pharmaceutical formulation) as a lyophilized powder and the other ofwhich contains a liquid for resuspending the formulation (e.g.,pharmaceutical formulation). In an embodiment, the kit further includesinstructions for use of the formulation. The kit may contain a notice asrequired by governmental agency regulating the manufacture, use, andsale of pharmaceuticals or biological products, the notice indicatingthat the formulation has been approved for manufacture, use, and/or salefor administration to humans. The formulation may be supplied in ahermetically-sealed container. The formulation may be provided as aliquid or as a lyophilized powder that can be reconstituted by theaddition, e.g., of water or saline, to a concentration suitable foradministration to a subject.

The kit optionally includes a device suitable for administering theformulation, e.g., a syringe or device for delivering particles oraerosols, e.g., an inhaler, a spray device, or a dropper or othersuitable delivery device. The device can be provided pre-loaded with oneor both of the agents or can be empty but suitable for loading.

Diagnostic Methods

The binding agents, e.g., antibody molecules, provided herein are usefulfor identifying the presence of influenza in a biological sample, e.g.,a patient sample, such as a fluid sample, e.g., a blood, serum, saliva,mucous, or urine sample, or a tissue sample, such as a biopsy.

In an embodiment, a patient sample is contacted with a binding agent,e.g., an antibody molecule, disclosed herein, and binding is detected.Binding can be detected with a number of formats and means of detection,e.g., with an antigen capture assay, such as an ELISA assay or Westernblot, or an immunohistochemistry assay. In an embodiment, the bindingagent, e.g., an antibody molecule, is provided, e.g., coupled to aninsoluble matrix, e.g., a bead or other substrate, and a detectionmolecule used to detect binding of HA.

Binding of binding agent, e.g., antibody molecule, to HA, can bedetected with a reagent comprising a detectable moiety, e.g., a reagent,e.g., an antibody, which binds the binding agent, e.g., antibodymolecule. In an embodiment, the binding agent, e.g., antibody molecule,has a detectable moiety. Suitable detectable moieties include enzymes(e.g., horseradish peroxidase, beta-galactosidase, luciferase, alkalinephosphatase, acetylcholinesterase, glucose oxidase and the like),radiolabels (e.g., ³H, ¹⁴C, ¹⁵N, ³⁵S, ⁹⁰Y, ⁹⁹Tc, ¹¹¹In, ¹²⁵I, ¹³¹I),haptens, fluorescent labels (e.g., FITC, rhodamine, lanthanidephosphors, fluorescein, fluorescein isothiocyanate, rhodamine,5-dimethylamine-1-napthalenesulfonyl chloride, phycoerythrin and thelike), phosphorescent molecules, chemiluminescent molecules,chromophores, luminescent molecules, photoaffinity molecules, coloredparticles or affinity ligands, such as biotin, predetermined polypeptideepitopes recognized by a secondary reporter (e.g., leucine zipper pairsequences, or binding sites for secondary antibodies, metal bindingdomains, epitope tags). In some embodiments, labels are attached byspacer arms of various lengths to reduce potential steric hindrance.

In an embodiment, a human is tested for presence of influenza virus be amethod described herein, and if the test is positive, a binding agents,e.g., antibody molecules, e.g., an antibody, provided herein, isadministered.

The binding agents, e.g., antibody molecules, e.g., an antibody,provided herein can be used for cytology assays, such as to identify anHA in a cell. The assay can be a colorimetric assay. A biological samplefrom a normal (non-infected) individual is used as a control. Thediagnostic assay can be performed in vitro.

The diagnostic assay can also be performed to determine infection ofcells in culture, e.g., of mammalian cells in culture. The antibodymolecules can be used in in vitro assays.

Because the antibody molecules disclosed herein bind a broad spectrum ofHA subtypes, the diagnostic assays disclosed herein can detect thepresence of influenza virus in patients infected with a variety ofdistinct strains of influenza. A patient sample can be further testedwith subtype specific antibodies, or other assays (e.g., RFLP(Restriction Fragment Length Polymorphism), PCR

(Polymerase Chain Reaction), RT-PCR (Reverse Transcription coupled toPolymerase Chain Reaction), Northern blot, Southern blot or DNAsequencing) to further determine the particular strain of virus.

In an embodiment, a patient determined to be infected with influenza Acan be further administered an antibody molecule disclosed herein, totreat the infection.

Also provided are solid substrates, e.g., beads, dipsticks, arrays, andthe like, on which is disposed a binding agent, e.g., antibody molecule.

The disclosure is further illustrated by the following examples, whichshould not be construed as further limiting.

Anti-HA antibody molecules described herein are also disclosed inInternational Publication No. WO2013/170139, U.S. Pat. No. 8,877,200,U.S. Pat. No. 9,096,657, and U.S. Patent Application Publication No. US2013/0302349. The contents of the aforesaid publications areincorporated by reference in their entirety.

TABLE 4C Nucleic acid and amino acid sequence SEQ ID Lab NO. no. SourceComment 1 n.a.  Table 2 Consensus AA sequence of HC CDR1 [S/T]Y[A/G]MH 2n.a. Table 2 Consensus AA sequence of HC CDR2V[I/V/L]S[Y/F]DG[S/N][Y/N][K/R]YYADSVQG 3 n.a. Table 2Consensus AA sequence of HC CDR3D[S/T][R/K/Q]LR[S/T]LLYFEWLS[Q/S]G[Y/L/V][F/L][N/D] 4 n.a. Table 2Consensus AA sequence of LC CDR1 Q[S/T][V/L/I][T/S][Y/F/W][N/S/D]YKNYLA170 n.a. Table 2 Consensus AA sequence of LC CDR1Q[S/T][V/L/I][T/S][Y/F/W][N/S/D/Q/R/E]YKNYLA 5 n.a. Table 2Consensus AA sequence of LC CDR2 W[A/G]S[T/A/Y/H/K/D][R/L]E[S/T] 6 n.a.Table 2 Consensus AA sequence of LC CDR3 QQ[Y/H]YRTPP[T/S] 7 n.a.Table 2 Consensus AA sequence of HC FR1[E/Q]VQLLE[S/T]GGGLVKPGQSLKLSCAASGFTF[S/T] 8 n.a. Table 2Consensus AA sequence of HC FR2 WVRQPPGKGLEWVA 9 n.a. Table 2Consensus AA sequence of HC FR3 RFTISRDNSKNTLYLQMNSLRAEDTAVYYCAK 10 n.a.Table 2 Consensus AA sequence of HC FR4 WG[A/Q]G[T/A][T/M][L/V]TVSS 11n.a. Table 2 Consensus AA sequence of LC FR1[E/D]I[V/Q]MTQSP[D/S][S/T][L/V][A/S][V/A][S/T][L/V/R]G[E/D]R[A/V][T/S]I[N/T/Q/D/R]C[K/R]SS 12 n.a. Table 2Consensus AA sequence of LC FR2 WYQQKPG[Q/K][P/A]PKLLIY 13 n.a. Table 2Consensus AA sequence of LC FR3GVP[D/E/S]RFSGSGSGTDFTLTISSLQ[A/P]ED[V/F/K/D]A[V/T]YYC 14 n.a. Table 2Consensus AA sequence of LC FR4 FG[G/Q/T/S/N]GTK[L/V][D/E]IK 15 15Table 3  AA sequence of HC VR of Ab A18; entireEVQLLESGGGLVKPGQSLKLSCAASGFTFTSYGMHWVRQPPGKGLEWVAVISYDGS VH15 TableHC domain is in FIG. 1; ID version is in FIG.YKYYADSVQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSRLRSLLYFEWLS 4A,5; NT sequence is in Example 1 QGYFNPWGAGTTLTVSS FIG. 2 28 28 Table 3,AA sequence of LC VR of Ab A18; entireEIVMTQSPDSLAVSLGERATINCKSSQSVTYNYKNYLAWYQQKPGQPPKLLIYWAS VL28 Table 4ALC domain is in FIG. 1; ID version is in FIG.TRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDIK FIG. 3A6A; NT sequence is in Example 1 16 16 Table 3AA sequence of HC VR of Abs 014, 028;EVQLLESGGGLVKPGQSLKLSCAASGFTFSSYGMHWVRQPPGKGLEWVAVVSYDGS VH16 Table 4AID version is in FIG. 5; NT sequence is inNKYYADSVQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDTKLRSLLYFEWLS FIG. 2Example 1 SGLLDYWGQGAMVTVSS 29 29 Table 3AA sequence of LC VR of Abs 014, 154,EIVMTQSPDSLAVSLGERATINCKSSQSVTFSYKNYLAWYQQKPGQPPKLLIYWAS VL29 Table 4A157; ID version is in FIG. 6A; NT sequenceTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDIK FIG. 3Ais in Example 1 30 30 Table 3 AA sequence of LC VR of Abs 028, 155;EIVMTQSPDSLAVSLGERATINCKSSQSVTFSYKNYLAWYQQKPGQPPKLLIYWAS VL30 Table 4AID version is in FIG. 6A; NT sequence is inTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDIK FIG. 3AExample 1 17 17 Table 3 AA sequence of HC VR of Abs 001, 009,EVQLLESGGGLVKPGQSLKLSCAASGFTFSSYGMHWVRQPPGKGLEWVAVVSYDGS VH17 Table 4A017, 025, 160, 186, 187, 188, 189, 190, 191,YKYYADSVQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSRLRSLLYFEWLS FIG. 2192, 193, 202, 211; ID version is in FIG. 5 QGYFNPWGAGTTLTVSS 31 31Table 3 AA sequence of LC VR of Abs 001, 002,EIVMTQSPDSLAVSLGERATINCKSSQSVTFSYKNYLAWYQQKPGQPPKLLIYWAS VL31 Table 4A003; ID version is in FIG. 6ATRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDIK FIG. 3A 18 18Table 3 AA sequence of HC VR of Abs 002, 010,EVQLLESGGGLVKPGQSLKLSCAASGFTFSSYGMHWVRQPPGKGLEWVAVVSYDGS VH18 Table 4AB18, 026, 203, 212; ID version is in FIG. 5YKYYADSVQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSRLRSLLYFEWLS FIG. 2QGYFNPWGAGTTLTVSS 19 19 Table 3 AA sequence of HC VR of Abs 003, 011,EVQLLESGGGLVKPGQSLKLSCAASGFTFSSYGMHWVRQPPGKGLEWVAVVSYDGS VH19 Table 4A019, 027, 194, 195, 196, 197, 198, 199, 200,YKYYADSVQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSRLRSLLYFEWLS FIG. 2204, 213; ID version is in FIG. 5 QGYFNPWGAGTTLTVSS 32 32 Table 3AA sequence of LC VR of Abs 009, 010,EIVMTQSPDSLAVSLGERATINCKSSQSVTFSYKNYLAWYQQKPGQPPKLLIYWAS VL32 Table 4A011; ID version is in FIG. 6ATRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDIK FIG. 3A 33 33Table 3 AA sequence of LC VR of Abs 017, B18,EIVMTQSPDSLAVSLGERATINCKSSQSVTFSYKNYLAWYQQKPGQPPKLLIYWAS VL33 Table 4A019; ID version is in FIG. 6ATRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDIK FIG. 3A 34 34Table 3 AA sequence of LC VR of Abs 025, 026,EIVMTQSPDSLAVSLGERATINCKSSQSVTFSYKNYLAWYQQKPGQPPKLLIYWAS VL34 Table 4A027, 086; ID version is in FIG. 6ATRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDIK FIG. 3A 20 20Table 3 AA sequence of HC VR of Ab 086; IDEVQLLESGGGLVKPGQSLKLSCAASGFTFSSYGMHWVRQPPGKGLEWVAVVSYDGS VH20 Table 4Aversion is in FIG. 5NRYYADSVQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSQLRSLLYFEWLS FIG. 2SGVLDYWGQGAMVTVSS 21 21 Table 3 AA sequence of HC VR of Abs 154,155; IDEVQLLESGGGLVKPGQSLKLSCAASGFTFSSYGMHWVRQPPGKGLEWVAVVSYDGN VH21 Table 4Aversion is in FIG. 5NKYYADSVQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSKLRSLLYFEWLS FIG. 2SGLLDYWGQGAMVTVSS 22 22 Table 3 AA sequence of HC VR of Abs 157, 159;EVQLLESGGGLVKPGQSLKLSCAASGFTFTTYAMHWVRQPPGKGLEWVAVVSYDGN VH22 Table 4AID version is in FIG. 5NKYYADSVQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSKLRSLLYFEWLS FIG. 2SGLLDYWGQGAMVTVSS 35 35 Table 3 AA sequence of LC VR of Ab 159; IDEIVMTQSPDSLAVSLGERATINCKSSQSVTFSYKNYLAWYQQKPGQPPKLLIYWAS VL35 Table 4Aversion is in FIG. 6ATRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDIK FIG. 3A 36 36Table 3 AA sequence of LC VR of Ab 160; IDEIVMSQSPDTLAVTLGERASINCKSSQTVTFNYKNYLAWYQQKPGQPPKVLIYWAS VL36 Table 4Aversion is in FIG. 6AARETGVPERFSGSGSGTDFTLTISSLQAEDVAVYYCQQHYRTPPSFGQGTKLEIK FIG. 3A 37 37Table 3 AA sequence of LC VR of Abs 186, 194;EIVMTQSPDSLAVSLGERATINCKSSQSVTFSYKNYLAWYQQKPGQPPKLLIYWAS VL37 Table 4AID version is in FIG. 6ATRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDIK 38 38 Table 3AA sequence of LC VR of Abs 187, 195;EIVMTQSPDSLAVSLGERATINCKSSQSVTFSYKNYLAWYQQKPGQPPKLLIYWAS VL38 Table 4AID version is in FIG. 6ATRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDIK FIG. 3A 39 39Table 3 AA sequence of LC VR of Abs 188, 196;EIVMTQSPDSLAVSLGERATINCKSSQSVTFSYKNYLAWYQQKPGQPPKLLIYWAS VL39 Table 4AID version is in FIG. 6ATRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDIK FIG. 3A 40 40Table 3 AA sequence of LC VR of Abs 189, 197;EIVMTQSPDSLAVSLGERATINCKSSQSVTFSYKNYLAWYQQKPGQPPKLLIYWAS VL40 Table 4AID version is in FIG. 6ATRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDIK FIG. 3A 41 41Table 3 AA sequence of LC VR of Abs 190, 198;EIVMTQSPDSLAVSLGERATINCKSSQSVTFSYKNYLAWYQQKPGQPPKLLIYWAS VL41 Table 4AID version is in FIG. 6ATRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDIK FIG. 3A 42 42Table 3 AA sequence of LC VR of Abs 191, 199;EIVMTQSPDSLAVSLGERATINCKSSQSVTFSYKNYLAWYQQKPGQPPKLLIYWAS VL42 Table 4AID version is in FIG. 6ATRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDIK FIG. 3A 43 43Table 3 AA sequence of LC VR of Abs 192, 200;EIVMTQSPDSLAVSLGERATINCKSSQSVTFSYKNYLAWYQQKPGQPPKLLIYWAS VL43 Table 4AID version is in FIG. 6ATRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDIK FIG. 3A 44 44Table 3 AA sequence of LC VR of Abs 193; IDEIVMTQSPDSLAVSLGERATINCKSSQSVTFSYKNYLAWYQQKPGQPPKLLIYWAS VL44 Table 4Aversion is in FIG. 6ATRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDIK FIG. 3A 45 45Table 3 AA sequence of LC VR of Abs 202, 203,DIQMTQSPSSLSASVGDRVTITCRSSQSITFNYKNYLAWYQQKPGKAPKLLIYWGS VL45 Table 4A204, 210, 031, 032, 033, 034; ID version isYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEIK FIG. 3Ain FIG. 6A; NT sequence is in Example 1 46 46 Table 3AA sequence of LC VR of Abs 211, 212,DIQMTQSPSSLSASVGDRVTITCRSSQSITFNYKNYLAWYQQKPGKAPKLLIYWGS VL46 Table 4A213, 219, 037, 038, 039, 040; ID version is YLESGVPSRFSGSGSGTDFTLTFIG. 3A in FIG. 6A 23 23 Table 3 AA sequence of HC VR of Abs 210, 219 ;EVQLLESGGGLVKPGQSLKLSCAASGFTFTSYGMHWVRQPPGKGLEWVAVVSYDGN VH23 Table 4AID version is in FIG. 5YKYYADSVQGFFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSKLRSLLYFEWLS FIG. 2QGYFNPWGAGTTLTVSS 24 24 Table 3 AA sequence of HC VR of Abs A001,EVQLLESGGGLVKPGQSLKLSCAASGFTFTSYGMHWVRQPPGKGLEWVAVVSYDGN VH24 Table 4AA002, A003, A010, A011, 031, 037; IDYKYYADSVQGFFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSKLRSLLYFEWLS FIG. 2version is in FIG. 5; NT sequence is in QGYFNPWGAGTTLTVSS Example 1 4747 Table 3 AA sequence of LC VR of Abs A001, 004,EIVMTQSPDSLAVSLGERATINCKSSQSVTFSYKNYLAWYQQKPGQPPKLLIYWAS VL47 Table 4A007, 016; ID version is in FIG. 6ATRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDIK FIG. 3A 48 48Table 3 AA sequence of LC VR of Abs 002, 005,EIVMTQSPDSLAVSLGERATINCKSSQSVTFSYKNYLAWYQQKPGQPPKLLIYWAS VL48 Table 4A008, A017; ID version is in FIG. 6ATRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDIK FIG. 3A 25 25Table 3 AA sequence of HC VR of Abs 004, 005,QVQLLETGGGLVKPGQSLKLSCAASGFTFTSYAMHWVRQPPGKGLEWVAVVSYDGN VH25 Table 4A006, 012, 013, 032, 038, 043, 044, 045, 046,YKYYADSVQGFFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSKLRSLLYFEWLS FIG. 2047, 048, 049, 050, 051, 052, 067, 068, 069, QGYFNPWGAGTTLTVSS070, 073, 074, 075, 076, 077; ID version isin FIG. 5; NT sequence is in Example 1 49 49 Table 3AA sequence of LC VR of Abs A003, 006,DIVMTQSPDTVAVTLGERATIDCKSSQTVTFNYKNYLAWYQQKPGQPPKLLIYWAS VL49 Table 4AA009, C18; ID version is in FIG. 6ATRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDIK FIG. 3A 26 26Table 3 AA sequence of HC VR of Abs 007, 008,EVQLLESGGGLVKPGQSLKLSCAASGFTFTSYAMHWVRQPPGKGLEWVAVVSYDGN VH26 Table 4AA009, A14, 015, 033, 039; ID version is inYKYYADSVQGFFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSKLRSLLYFEWLS FIG. 2 FIG. 5QGYFNPWGAGTTLTVSS 50 50 Table 3 AA sequence of LC VR of Abs A010 012,DIVMTQSPDTVAVTLGERATIDCKSSQTVTFNYKNYLAWYQQKPGQPPKLLIYWAS VL50 Table 4AA14, A019; ID version is in FIG. 6ATRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDIK FIG. 3A 51 51Table 3 AA sequence of LC VR of Ab YWASDIVMTQSPDTVAVTLGERATIDCKSSQTVTFNYKNYLAWYQQKPGQPPKLLIYWAS VL51 Table 4A015; ID version is in FIG. 6ATRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDIK FIG. 3A 27 27Table 3 AA sequence of HC VR of Abs 016, A017,EVQLLESGGGLVKPGQSLKLSCAASGFTFTSYAMHWVRQPPGKGLEWVAVVSYDGN VH27 Table 4AC18, A019, 034, 040; ID version is in FIG. 5YKYYADSVQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSRLRTLLYFEWLS FIG. 2QGYFNPWGAGTTLTVSS 60 60 Table 3 AA sequence of LC VR of Ab 043; IDDIQMTQSPSSLSASVGDRVTITCRSSQSITFNYKNYLAWYQQKPGKAPKLLIYWGS VL60 Table 4Aversion is in FIG. 6BYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEIK FIG. 3B 52 52Table 3 AA sequence of LC VR of Abs 044, 071,DIQMTQSPSSLSASVGDRVTITCRSSQSITFNYKNYLAWYQQKPGKAPKLLIYWGS VL52 Table 4A072, 078; ID version is in FIG. 6B; NTYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEIK FIG. 3Bsequence is in Example 1 57 57 Table 3AA sequence of LC VR of Ab 045; IDDIQMTQSPSSLSASVGDRVTITCRSSQSITFNYKNYLAWYQQKPGKAPKLLIYWGS VL57 Table 4Aversion is in FIG. 6BYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEIK FIG. 3B 59 59Table 3 AA sequence of LC VR of Ab 046; IDDIQMTQSPSSLSASVGDRVTITCRSSQSITFNYKNYLAWYQQKPGKAPKLLIYWGS VL59 Table 4Aversion is in FIG. 6BYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEIK FIG. 3B 55 55Table 3 AA sequence of LC VR of Ab 047; IDDIQMTQSPSSLSASVGDRVTITCRSSQSITFNYKNYLAWYQQKPGKAPKLLIYWGS VL55 Table 4Aversion is in FIG. 6BKLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEIK FIG. 3B 58 58Table 3 AA sequence of LC VR of Ab 048; IDDIQMTQSPSSLSASVGDRVTITCRSSQSITFNYKNYLAWYQQKPGKAPKLLIYWGS VL58 Table 4Aversion is in FIG. 6BYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEIK FIG. 3B 54 54Table 3 AA sequence of LC VR of Ab 049; IDDIQMTQSPSSLSASVGDRVTITCRSSQSITFNYKNYLAWYQQKPGKAPKLLIYWGS VL54 Table 4Aversion is in FIG. 6BHLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEIK FIG. 3B 56 56Table 3 AA sequence of LC VR of Ab 050; IDDIQMTQSPSSLSASVGDRVTITCRSSQSITFNYKNYLAWYQQKPGKAPKLLIYWGS VL56 Table 4Aversion is in FIG. 6BDLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEIK 53 53 Table 3AA sequence of LC VR of Ab 051; IDDIQMTQSPSSLSASVGDRVTITCRSSQSITFNYKNYLAWYQQKPGKAPKLLIYWGS VL53 Table 4Aversion is in FIG. 6BTLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEIK FIG. 3B 61 61Table 3 AA sequence of LC VR of Ab 052; IDDIVMTQSPDTVAVTLGERATIDCKSSQTVTFNYKNYLAWYQQKPGQPPKLLIYWAS VL61 Table 4Aversion is in FIG. 6BTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDIK FIG. 3B 153 153Table 3 AA sequence of LC VR of Ab 067; IDDIQMTQSPSSLSASVGDRVTITCRSSQSITFNYKNYLAWYQQKPGKAPKLLIYWGS VL153 Table 4Aversion is in FIG. 6BYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEIK FIG. 3B 154 154Table 3 AA sequence of LC VR of Ab 068; IDDIQMTQSPSSLSASVGDRVTITCRSSQSITFNYKNYLAWYQQKPGKAPKLLIYWGS VL154 Table 4Aversion is in FIG. 6BYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEIK FIG. 3B 155 155Table 3 AA sequence of LC VR of Abs 069, 079;DIQMTQSPSSLSASVGDRVTITCRSSQSITFEYKNYLAWYQQKPGKAPKLLIYWGS VL155 Table 4AID version is in FIG. 6BYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEIK FIG. 3B 156 156Table 3 AA sequence of LC VR of Ab 070; IDDIVMTQSPDTVAVTLGERATIDCKSSQTVTFNYKNYLAWYQQKPGQPPKLLIYWAS VL156 Table 4Aversion is in FIG. 6BTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDIK FIG. 3B 162 162Table 3 AA sequence of HC VR of Ab 071EVQLLESGGGLVKPGQSLKLSCAASGFTFTSYAMHWVRQPPGKGLEWVAVVSYDGN VL162 Table 4AYKYYADSVQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSRLRTLLYFEWLS FIG. 7QGYFNPWGQGTTLTVSS 163 163 Table 3 AA sequence of HC VR of Ab 072EVQLLESGGGLVKPGQSLKLSCAASGFTFTSYAMHWVRQPPGKGLEWVAVVSYDGN VL163 Table 4AYKYYADSVQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSRLRTLLYFEWLS FIG. 7QGYFNPWGQGTTLTVSS 165 165 Table 3 AA sequence of LC VR of Ab 073DIQMTQSPSSLSASVGDRVTITCRSSQSITFEYKNYLAWYQQKPGKAPKLLIYWGS VL165 Table 4AYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEIK FIG. 7 166 166Table 3 AA sequence of LC VR of Abs 074, 080DIQMTQSPSSLSASVGDRVTITCRSSQSITFEYKNYLAWYQQKPGKAPKLLIYWGS VL166 Table 4AYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEIK FIG. 7 167 167Table 3 AA sequence of LC VR of Ab 075DIQMTQSPSSLSASVGDRVTITCRSSQSITFEYKNYLAWYQQKPGKAPKLLIYWGS VL167 Table 4AYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEIK FIG. 7 168 168Table 3 AA sequence of LC VR of Ab 076DIQMTQSPSSLSASVGDRVTITCRSSQSITFEYKNYLAWYQQKPGKAPKLLIYWGS VL168 Table 4AYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEIK FIG. 7 169 169Table 3 AA sequence of LC VR of Abs 077, 081DIQMTQSPSSLSASVGDRVTITCRSSQSITFEYKNYLAWYQQKPGKAPKLLIYWGS VL169 Table 4AYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEIK FIG. 7 164 164Table 3 AA sequence of HC VR of Abs 078, 079,QVQLLETGGGLVKPGQSLKLSCAASGFTFTSYAMHWVRQPPGKGLEWVAVVSYDGN VL164 Table 4A080, 081 YKYYADSVQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSRLRTLLYFEWLS FIG. 7QGYFNPWGQGTTLTVSS 161 HC161 Table 4A AA sequence of HC VR consensus; IDEVQLLESGGGLVKPGQSLKLSCAASGFTFSSYGMHWVRQPPGKGLEWVAVVSYDGS FIG. 2version is in FIG. 5NKYYADSVQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSKLRSLLYFEWLSSGLLDYWGQGAMVTVSS 62 LC62 Table 4A AA sequence of LC VR consensus; IDDIQMTQSPSSLSASVGDRVTITCRSSQSITFEYKNYLAWYQQKPGKAPKLLIYWGS FIG. 3Bversion is in FIG. 6BYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEIK 96 15-IDTable 4B AA sequence of HC VR of Ab A18; non-IDIDEVQLLESGGGLVKPGQSLKLSCAASGFTFTSYGMHWVRQPPGKGLEWVAVISYD FIG. 5version is in FIG. 2GSYKYYADSVQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSRLRSLLYFEWLSQGYFNPWGAGTTLTVSS 110 28-ID Table 4BAA sequence of LC VR of Ab A18; non-IDIDEVQLLESGGGLVKPGQSLKLSCAASGFTFTSYGMHWVRQPPGKGLEWVAVISYD FIG. 6Aversion is in FIGS. 3A-3BASTESDGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDI K 97 16-IDTable 4B AA sequence of HC VR of Abs 014, 028;IDEVQLLESGGGLVKPGQSLKLSCAASGFTFTSYGMHWVRQPPGKGLEWVAVISYD FIG. 5non-ID version is in FIG. 2GSYKYYADSVQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSRLRSLLYFEWLSQGYFNPWGAGTTLTVSS 111 29-ID Table 4BAA sequence of LC VR of Abs 014, 154,IDEVQLLESGGGLVKPGQSLKLSCAASGFTFTSYGMHWVRQPPGKGLEWVAVISYD FIG. 6A157; non-ID version is in FIGS. 3A-3BASTESDGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDI K 98 17-IDTable 4B AA sequence of HC VR of Ab 001, 009,IDEVQLLESGGGLVKPGQSLKLSCAASGFTFTSYGMHWVRQPPGKGLEWVAVISYD FIG. 5017, 025, 160, 186, 187, 188, 189, 190, 191,GNYKYYADSVQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSRLRSLLYFEW192, 193, 202, 211; non-ID version is in LSQGYFNPWGAGTTLTVSS FIG. 2 11230-ID Table 4B AA sequence of LC VR of Abs 028, 155;IDEVQLLESGGGLVKPGQSLKLSCAASGFTFTSYGMHWVRQPPGKGLEWVAVISYD FIG. 6Anon-ID version is in FIGS. 3A-3B IASTESDGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDI K 99 18-IDTable 4B AA sequence of HC VR of Abs 002, 010,IDEVQLLESGGGLVKPGQSLKLSCAASGFTFTSYGMHWVRQPPGKGLEWVAVISYD FIG. 5B18, 026, 203, 212; non-ID version is inGNYKYYADSVQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSRLRSLLYFEW FIG. 2LSQGYFNPWGAGTTLTVSS 113 35-ID Table 4BAA sequence of LC VR of Ab 159; non-IDIDEVQLLESGGGLVKPGQSLKLSCAASGFTFTSYGMHWVRQPPGKGLEWVAVISYD FIG. 6Aversion is in FIGS. 3A-3BASTESDGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDI K 100 19-IDTable 4B AA sequence of HC VR of Abs 003, 011,IDEVQLLESGGGLVKPGQSLKLSCAASGFTFTSYGMHWVRQPPGKGLEWVAVISYD FIG. 5019, 027, 194, 195, 196, 197, 198, 199, 200,GNYKYYADSVQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSRLRSLLYFEW204, 213; non-ID version is in FIG. 2 LSQGYFNPWGAGTTLTVSS 114 31-IDTable 4B AA sequence of LC VR of Abs 001, 002,IDEVQLLESGGGLVKPGQSLKLSCAASGFTFTSYGMHWVRQPPGKGLEWVAVISYD FIG. 6A003; non-ID version is in FIGS. 3A-3BASTESDGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDI K 101 21-IDTable 4B AA sequence of HC VR of Abs 154,155;IDEVQLLESGGGLVKPGQSLKLSCAASGFTFTSYGMHWVRQPPGKGLEWVAVISYD FIG. 5non-ID version is in FIG. 2GNYKYYADSVQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSRLRSLLYFEWLSSGLLDYWGQGAMVTVSS 115 32-ID Table 4BAA sequence of LC VR of Abs 009, 010,IDEVQLLESGGGLVKPGQSLKLSCAASGFTFTSYGMHWVRQPPGKGLEWVAVISYD FIG. 6A011; non-ID version is in FIGS. 3A-3BASTESDGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDI K 102 22-IDTable 4B AA sequence of HC VR of Abs 157, 159;IDEVQLLESGGGLVKPGQSLKLSCAASGFTFTSYGMHWVRQPPGKGLEWVAVISYD FIG. 5non-ID version is in FIG. 2GNYKYYADSVQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSRLRSLLYFEWLSSGLLDYWGQGAMVTVSS 116 33-ID Table 4BAA sequence of LC VR of Abs 017, B18,IDEVQLLESGGGLVKPGQSLKLSCAASGFTFTSYGMHWVRQPPGKGLEWVAVISYD FIG. 6A019; non-ID version is in FIGS. 3A-3BASTESDGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDI K 103 20-IDTable 4B AA sequence of HC VR of Ab 086; non-IDIDEVQLLESGGGLVKPGQSLKLSCAASGFTFTSYGMHWVRQPPGKGLEWVAVISYD FIG. 5version is in FIG. 2GNNKYYADSVQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSRLRSLLYFEWLSSGLLDYWGQGAMVTVSS 117 34-ID Table 4BAA sequence of LC VR of Abs 025, 026,IDEVQLLESGGGLVKPGQSLKLSCAASGFTFTSYGMHWVRQPPGKGLEWVAVISYD FIG. 6A027, 086; non-ID version is in FIGS. 3A-3BASTESDGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDI K 104 23-IDTable 4B AA sequence of HC VR of Abs 210, 219;IDEVQLLESGGGLVKPGQSLKLSCAASGFTFTSYGMHWVRQPPGKGLEWVAVISYD FIG. 5non-ID version is in FIG. 2GNYKYYADSVQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSRLRSLLYFEWLSQGLLDYWGQGAMVTVSS 118 36-ID Table 4BAA sequence of LC VR of Ab 160; non-IDIDEVQLLESGGGLVKPGQSLKLSCAASGFTFTSYGMHWVRQPPGKGLEWVAVISYD FIG. 6Aversion is in FIGS. 3A-3BASTESDGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDI K 105 24-IDTable 4B AA sequence of HC VR of Abs A001,IDEVQLLESGGGLVKPGQSLKLSCAASGFTFTSYGMHWVRQPPGKGLEWVAVISYD FIG. 5A002, A003, A010, A011, 031, 037; non-GNYKYYADSVQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSRLRSLLYFEWID version is in FIG. 2 LSQGLLDYWGQGAMVTVSS 119 45-ID Table 4BAA sequence of LC VR of Abs 202, 203,IDDIQMTQSPSSLSASVGDRVTITCRSSQSITFNYKNYLAWYQQKPGKAPKLLIYW FIG. 6A204, 210, 031, 032, 033, 034 ; non-IDGSYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEIversion is in FIGS. 3A-3B K 106 25-ID Table 4BAA sequence of HC VR of Abs 004, 005,IDQVQLLESGGGLVKPGQSLKLSCAASGFTFTSYGMHWVRQPPGKGLEWVAVISYD FIG. 5006, 012, 013, 032, 038, 043, 044, 045, 046,GNYKYYADSVQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSRLRSLLYFEW047, 048, 049, 050, 051, 052, 067, 068, 069, LSQGYFNPWGQGTTLTVSS070, 073, 074, 075, 076, 077; non-ID version is in FIG. 2 120 46-IDTable 4B AA sequence of LC VR of Abs 211, 212,IDDIQMTQSPSSLSASVGDRVTITCRSSQSITFNYKNYLAWYQQKPGKAPKLLIYW FIG. 6A213, 219, 037, 038, 039, 040; non-IDGSYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEIversion is in FIGS. 3A-3B K 107 26-ID Table 4BAA sequence of HC VR of Abs 007, 008,IDEVQLLESGGGLVKPGQSLKLSCAASGFTFTSYGMHWVRQPPGKGLEWVAVISYD FIG. 5A009, A14, 015, 033, 039; non-ID versionGNYKYYADSVQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSRLRSLLYFEW is in FIG. 2LSQGLLDYWGQGAMVTVSS 121 37-ID Table 4BAA sequence of LC VR of Abs 186, 194;IDEIMVTQSPDSLAVSLGERATINCKSSQTVTFNYKNYLAWYQQKPGQPPKLLIYW FIG. 6Anon-ID version is in FIGS. 3A-3BASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQHYRTPPSFGTGTKLDI K 108 27-IDTable 4B AA sequence of HC VR of Abs 016, A017,IDEVQLLESGGGLVKPGQSLKLSCAASGFTFTSYGMHWVRQPPGKGLEWVAVISYD FIG. 5C18, A019, 034, 040; non-ID version is inGNYKYYADSVQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSRLRSLLYFEW FIG. 2LSQGLLDYWGQGAMVTVSS 122 38-ID Table 4BAA sequence of LC VR of Abs 187, 195;IDEIMVTQSPDSLAVSLGERATINCKSSQTVTFNYKNYLAWYQQKPGQPPKLLIYW FIG. 6Anon-ID version is in FIGS. 3A-3BASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQHYRTPPSFGTGTKLDI K 109 161-IDTable 4B AA sequence of HC VR consensus ID; non-IDEVQLLESGGGLVKPGQSLKLSCAASGFTFTSYGMHWVRQPPGKGLEWVAVISYD FIG. 5ID version is in FIG. 2GSNKYYADSVQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSRLRSLLYFEWLSSGLLDYWGQGAMVTVSS 123 39-ID Table 4BAA sequence of LC VR of Abs 188, 196;IDEIMVTQSPDSLAVSLGERATINCKSSQTVTFNYKNYLAWYQQKPGQPPKLLIYW FIG. 6Anon-ID version is in FIGS. 3A-3BASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQHYRTPPSFGTGTKLDI K 124 40-IDTable 4B AA sequence of LC VR of Abs 189, 197;IDEIMVTQSPDSLAVSLGERATINCKSSQTVTFNYKNYLAWYQQKPGQPPKLLIYW FIG. 6Anon-ID version is in FIGS. 3A-3BASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQHYRTPPSFGTGTKLDI K 125 41-IDTable 4B AA sequence of LC VR of Abs 190, 198;IDEIMVTQSPDSLAVSLGERATINCKSSQTVTFNYKNYLAWYQQKPGQPPKLLIYW FIG. 6Anon-ID version is in FIGS. 3A-3BASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQHYRTPPSFGTGTKLDI K 126 42-IDTable 4B AA sequence of LC VR of Abs 191, 199;IDEIMVTQSPDSLAVSLGERATINCKSSQTVTFNYKNYLAWYQQKPGQPPKLLIYW FIG. 6Anon-ID version is in FIGS. 3A-3BASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQHYRTPPSFGTGTKLDI K 127 43-IDTable 4B AA sequence of LC VR of Abs 192, 200;IDEIMVTQSPDSLAVSLGERATINCKSSQTVTFNYKNYLAWYQQKPGQPPKLLIYW FIG. 6Anon-ID version is in FIGS. 3A-3BASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQHYRTPPSFGTGTKLDI K 128 44-IDTable 4B AA sequence of LC VR of Abs 193; non-IDIDEIMVTQSPDSLAVSLGERATINCKSSQTVTFNYKNYLAWYQQKPGQPPKLLIYW FIG. 6Aversion is in FIGS. 3A-3BASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQHYRTPPSFGTGTKLDI K 129 47-IDTable 4B AA sequence of LC VR of Abs A001, 004,IDDIMVTQSPDSLAVSLGERATINCKSSQTVTFNYKNYLAWYQQKPGQPPKLLIYW FIG. 6A007, 016 ASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQHYRTPPSFGTGTKLDI K 13048-ID Table 4B AA sequence of LC VR of Abs 002, 005,IDDIMVTQSPDSLAVSLGERATINCKSSQTVTFNYKNYLAWYQQKPGQPPKLLIYW FIG. 6A008, A017; non-ID version is in FIGS. 3A-ASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQHYRTPPSFGTGTKLDI 3B K 131 49-IDTable 4B AA sequence of LC VR of Abs A003, 006,IDDIMVTQSPDSLAVSLGERATINCKSSQTVTFNYKNYLAWYQQKPGQPPKLLIYW FIG. 6AA009, C18; non-ID version is in FIGS. 3A-ASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQHYRTPPSFGTGTKLDI 3B K 132 50-IDTable 4B AA sequence of LC VR of Abs A010 012,IDDIMVTQSPDSLAVSLGERATINCKSSQTVTFNYKNYLAWYQQKPGQPPKLLIYW FIG. 6AA14, A019; non-ID version is in FIGS. 3A-ASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQHYRTPPSFGTGTKLDI 3B K 133 51-IDTable 4B AA sequence of LC VR of Ab A011, 013,IDDIMVTQSPDSLAVSLGERATINCKSSQTVTFNYKNYLAWYQQKPGQPPKLLIYW FIG. 6A015; non-ID version is in FIGS. 3A-3BASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQHYRTPPSFGTGTKLDI K 134 52-IDTable 4B AA sequence of LC VR of Abs 044, 071,IDDIQMTQSPSSLSASVGDRVTITCRSSQSITFNYKNYLAWYQQKPGKAPKLLIYW FIG. 6B072, 078; non-ID version is in FIGS. 3A-3BGSYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEI K 135 53-IDTable 4B AA sequence of LC VR of Ab 051; non-IDIDDIQMTQSPSSLSASVGDRVTITCRSSQSITFNYKNYLAWYQQKPGKAPKLLIYW FIG. 6Bversion is in FIGS. 3A-3BGSYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEI K 136 54-IDTable 4B AA sequence of LC VR of Ab 049; non-IDIDDIQMTQSPSSLSASVGDRVTITCRSSQSITFNYKNYLAWYQQKPGKAPKLLIYW FIG. 6Bversion is in FIGS. 3A-3BGSYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEI K 137 55-IDTable 4B AA sequence of LC VR of Ab 047; non-IDIDDIQMTQSPSSLSASVGDRVTITCRSSQSITFNYKNYLAWYQQKPGKAPKLLIYW FIG. 6Bversion is in FIGS. 3A-3BGSYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEI K 138 56-IDTable 4B AA sequence of LC VR of Ab 050; non-IDIDDIQMTQSPSSLSASVGDRVTITCRSSQSITFNYKNYLAWYQQKPGKAPKLLIYW FIG. 6Bversion is in FIGS. 3A-3BGSYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEI K 139 57-IDTable 4B AA sequence of LC VR of Ab 045; non-IDIDDIQMTQSPSSLSASVGDRVTITCRSSQSITFNYKNYLAWYQQKPGKAPKLLIYW FIG. 6Bversion is in FIGS. 3A-3BGSYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEI K 140 58-IDTable 4B AA sequence of LC VR of Ab 048; non-IDIDDIQMTQSPSSLSASVGDRVTITCRSSQSITFNYKNYLAWYQQKPGKAPKLLIYW FIG. 6Bversion is in FIGS. 3A-3BGSYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEI K 141 59-IDTable 4B AA sequence of LC VR of Ab 046; non-IDIDDIQMTQSPSSLSASVGDRVTITCRSSQSITFNYKNYLAWYQQKPGKAPKLLIYW FIG. 6Bversion is in FIGS. 3A-3BGSYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEI K 142 60-IDTable 4B AA sequence of LC VR of Ab 043; non-IDIDDIQMTQSPSSLSASVGDRVTITCRSSQSITFNYKNYLAWYQQKPGKAPKLLIYW FIG. 6Bversion is in FIGS. 3A-3BGSYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEI K 143 61-IDTable 4B AA sequence of LC VR of Ab 052; non-IDIDDIQMTQSPSSLSASVGDRVTITCRSSQSITFNYKNYLAWYQQKPGKAPKLLIYW FIG. 6Bversion is in FIGS. 3A-3BGSYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEI K 157 153-IDTable 4B AA sequence of LC VR of Ab 067; non-IDIDDIQMTQSPSSLSASVGDRVTITCRSSQSITFNYKNYLAWYQQKPGKAPKLLIYW FIG. 6Bversion is in FIGS. 3A-3BGSYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEI K 158 154-IDTable 4B AA sequence of LC VR of Ab 068; non-IDIDDIQMTQSPSSLSASVGDRVTITCRSSQSITFNYKNYLAWYQQKPGKAPKLLIYW FIG. 6Bversion is in FIGS. 3A-3BGSYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEI K 159 155-IDTable 4B AA sequence of LC VR of Abs 069, 079;IDDIQMTQSPSSLSASVGDRVTITCRSSQSITFNYKNYLAWYQQKPGKAPKLLIYW FIG. 6Bnon-ID version is in FIGS. 3A-3BGSYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEI K 160 156-IDTable 4B AA sequence of LC VR of Ab 070; non-IDIDDIQMTQSPSSLSASVGDRVTITCRSSQSITFNYKNYLAWYQQKPGKAPKLLIYW FIG. 6Bversion is in FIGS. 3A-3BGSYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEI K 144 62-IDTable 4B AA sequence of LC VR consensus ID; non-IDDIQMTQSPSSLSASVGDRVTITCRSSQSITFNYKNYLAWYQQKPGKAPKLLIYW FIG. 6BID version is in FIGS. 3A-3BGSYLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYRTPPSFGQGTKVEI K 63 VH16Example NT sequence of HC VR of Abs 014, 028GAGGTACAGCTCCTCGAATCGGGAGGGGGACTGGTCAAACCCGGTCAATCGCTCAA 1ACTCTCGTGTGCAGCGTCAGGTTTTACGTTCAGCTCATATGGGATGCACTGGGTCCGCCAGCCTCCGGGAAAGGGACTGGAGTGGGTGGCAGTCGTGCGTATGACGGGAGCAATAAGTACTACGCCGATTCAGTGCAAGGTCGGTTTACCATTTCGAGGGATAACAGCAAGAACACGCTCTACTTGCAGATGAACTCACTTAGAGCGGAAGATACGGCTGTGTACTATTGCGCCAAAGACACAAAGCTGCGATCCCTGTTGTACTTCGAATGGTTGTCCTCGGGCTTGCTTGACTATTGGGGGCAGGGCGCCATGGTCACAGTATCCAGCGCGTC GACTAAGGGGCCC64 VL29 Example NT sequence of LC VR of Abs 014, 154,GAGATCGTGATGACGCAGAGCCCCGATAGCCTCGCTGTCTCATTGGGGGAACGGGC 1 157CACGATTAACTGCAAATCCTCACAGTCGGTGACTTTCAGCTATAAGAATTACCTGGCATGGTATCAGCAGAAGCCGGGTCAACCCCCAAAACTGTTGATCTACTGGGCCTCCACACGCGAGTCGGGAGTCCCGGACCGATTTTCGGGTTCAGGGTCCGGCACTGACTTTACCCTCACAATTTCATCGCTTCAAGCGGAGGATGTAGCAGTGTACTATTGTCAGCAGTATTACAGAACACCTCCCACCTTCGGAGGGGGAACGAAACTTGACATCAAGGGA TCC 65 VL30Example NT sequence of LC VR of Abs 028, 155GAGATCGTGATGACGCAGAGCCCCGATAGCCTCGCTGTCTCATTGGGGGAACGGGC 1CACGATTAACTGCAAATCCTCACAGTCGGTGACTTTCGACTATAAGAATTACCTGGCATGGTATCAGCAGAAGCCGGGTCAACCCCCAAAACTGTTGATCTACTGGGCCTCCACACGCGAGTCGGGAGTCCCGGACCGATTTTCGGGTTCAGGGTCCGGCACTGACTTTACCCTCACAATTTCATCGCTTCAAGCGGAGGATGTAGCAGTGTACTATTGTCAGCAGTATTACAGAACACCTCCCACCTTCGGAGGGGGAACGAAACTTGACATCAAGGGA TCC 66 VH15Example NT sequence of HC VR of Ab A18GAAGTGCAACTCCTCGAGTCAGGAGGAGGTTTGGTGAAACCGGGTCAGTCCTTGAA 1ACTGAGCTGTGCAGCAAGCGGGTTCACGTTTACGTCGTACGGCATGCACTGGGTACGGCAGCCTCCCGGGAAGGGACTTGAATGGGTCGCCGTCATCTCATACGACGGGTCGTACAAATACTATGCGGATAGCGTGCAAGGTCGCTTCACAATTTCCCGGGACAATTCGAAGAATACACTGTATCTTCAGATGAACTCGCTCAGGGCTGAGGACACGGCGGTCTATTACTGCGCGAAGGATTCGCGACTCAGATCCCTTTTGTACTTTGAGTGGCTGTCGCAGGGGTATTTCAACCCATGGGGAGCCGGAACCACTTTGACCGTATCAAGCGCGTC AACAAAGGGGCCC187 V128 Example NT sequence of LC VR of Ab A18GAAATTGTAATGACGCAGAGCCCTGATAGCCTTGCCGTGTCCCTGGGTGAGAGGGC 1GACAATCAATTGTAAGTCATCACAGTCGGTCACGTACAACTACAAGAACTACCTGGCGTGGTATCAACAGAAACCCGGGCAGCCGCCCAAATTGCTCATCTATTGGGCTTCGACACGGGAGTCGGGTGTGCCAGACCGCTTCTCCGGGTCAGGATCGGGAACTGACTTCACGTTGACTATTTCGTCCCTCCAGGCAGAAGATGTAGCCGTCTACTATTGCCAACAGTATTACAGAACGCCGCCTACATTTGGAGGCGGGACCAAACTTGACATCAAGGGATCCGTGGCCGCCCCCAGCGTCTTCATCTTCCCGCCCAGCGACGAGCAGCTGAAGTCGGGCACGGCCAGCGTGGTGTGCCTCCTGAACAACTTCTACCCCCGCGAGGCGAAGGTCCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGGAACAGCCAGGAGAGCGTGACCGAGCAGGACTCGAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAGGCCGACTACGAGAAGCACAAGGTCTACGCCTGCGAGGTGACCCACCAGGGGCTCTCGAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTG 149 VL52 ExampleNT sequence of LC VR of Abs 044, 071,GACATTCAGATGACTCAGTCGCCTTCGTCATTGTCCGCCTCCGTGGGTGATAGGGT 1 072, 078CACGATCACGTGCCGGAGCAGCCAGTCCATCACCTTCAATTACAAAAACTATTTGGCATGGTATCAACAGAAACCCGGAAAGGCGCCGAAGCTCCTGATCTACTGGGGTTCATATCTTGAGTCGGGGGTGCCGTCGAGATTTTCGGGCAGCGGATCAGGGACGGATTTCACGCTGACCATTTCGTCACTCCAGCCCGAGGACTTTGCGACATATTACTGTCAACAGCACTACAGGACACCCCCATCTTTCGGACAGGGGACTAAAGTAGAAATCAAGGGATCCGTGGCCGCCCCCAGCGTCTTCATCTTCCCGCCCAGCGACGAGCAGCTGAAGTCGGGCACGGCCAGCGTGGTGTGCCTCCTGAACAACTTCTACCCCCGCGAGGCGAAGGTCCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGGAACAGCCAGGAGAGCGTGACCGAGCAGGACTCGAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAGGCCGACTACGAGAAGCACAAGGTCTACGCCTGCGAGGTGACCCACCAGGGGCTCTCGAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCTGA 150 VL45 ExampleNT sequence of LC VR of Abs 202, 203,GACATTCAGATGACTCAGTCGCCTTCGTCATTGTCCGCCTCCGTGGGTGATAGGGT 1204, 210, 031, 032, 033, 034CACGATCACGTGCCGGAGCAGCCAGTCCATCACCTTCAATTACAAAAACTATTTGGCATGGTATCAACAGAAACCCGGAAAGGCGCCGAAGCTCCTGATCTACTGGGGTTCATATCTTGAGTCGGGGGTGCCGTCGAGATTTTCGGGCAGCGGATCAGGGACGGATTTCACGCTGACCATTTCGTCACTCCAGCCCGAGGACTTTGCGACATATTACTGTCAACAGCACTACAGGACACCCCCATCTTTCGGACAGGGGACTAAAGTAGAAATCAAGGGATCCGTGGCCGCCCCCAGCGTCTTCATCTTCCCGCCCAGCGACGAGCAGCTGAAGTCGGGCACGGCCAGCGTGGTGTGCCTCCTGAACAACTTCTACCCCCGCGAGGCGAAGGTCCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGGAACAGCCAGGAGAGCGTGACCGAGCAGGACTCGAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAGGCCGACTACGAGAAGCACAAGGTCTACGCCTGCGAGGTGACCCACCAGGGGCTCTCGAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCTGAGAATTC 151 VH25 Exampleyr sequence of FR: VIZ of Abs 004, 005,CAGGTACAATTGCTTGAGACAGGTGGAGGACTCGTGAAGCCAGGTCAGTCATTGAA 1006, 012, 013, 032, 038, 043, 044, 045, 046,ACTGAGCTGTGCCGCATCCGGGTTCACATTCACTTCCTACGCGATGCACTGGGTCC047, 048, 049, 050, 051, 052, 067, 068, 069,GCCAGCCTCCCGGAAAGGGACTTGAGTGGGTCGCTGTGGTATCGTATGATGGGAAT070, 073, 074, 075, 076, 077TACAAATACTATGCAGACTCCGTGCAAGGCCGGTTTACGATTAGCAGGGACAACTCGAAGAATACCCTTTACCTCCAAATGAACTCGCTCCGAGCGGAGGACACGGCGGTGTATTACTGCGCGAAGGATTCACGGTTGAGATCGCTGCTCTATTTTGAATGGTTGTCACAGGGGTACTTCAACCCGTGGGGTCAGGGAACAACACTGACCGTCAGCTCAGCCTCGACTAAAGGGCCCAGCGTGTTCCCGCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGGACCGCCGCCCTGGGCTGCCTCGTCAAGGACTACTTCCCCGAGCCCGTGACCGTGTCGTGGAACAGCGGCGCGCTGACGAGCGGGGTCCACACCTTCCCGGCCGTGCTGCAGAGCAGCGGCCTCTACTCGCTGAGCAGCGTGGTCACCGTGCCCAGCAGCAGCCTGGGGACCCAGACGTACATCTGCAACGTGAACCACAAGCCCTCGAACACCAAGGTCGACAAGAAGGTGGAGCCCCCGAAGAGCTGCGACAAAACTCACACATGCCCACCGTGCCCAGGTACTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGTGAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA   152 VH24 ExampleNT sequence of FR: VIZ of Abs A001,GAAGTACAATTGCTTGAGTCGGGTGGAGGACTCGTGAAGCCAGGTCAGTCATTGAA 1A002, A003, A010, A011, 031, 037ACTGAGCTGTGCCGCATCCGGGTTCACATTCACTTCCTACGCGATGCACTGGGTCCGCCAGCCTCCCGGAAAGGGACTTGAGTGGGTCGCTGTGGTATCGTATGATGGGAATTACAAATACTATGCAGACTCCGTGCAAGGCCGGTTTACGATTAGCAGGGACAACTCGAAGAATACCCTTTACCTCCAAATGAACTCGCTCCGAGCGGAGGACACGGCGGTGTATTACTGCGCGAAGGATTCACGGTTGAGATCGCTGCTCTATTTTGAATGGTTGTCACAGGGGTACTTCAACCCGTGGGGTCAGGGAACAACACTGACCGTCAGCTCAGCCTCGACTAAAGGGCCCAGCGTGTTCCCGCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGGACCGCCGCCCTGGGCTGCCTCGTCAAGGACTACTTCCCCGAGCCCGTGACCGTGTCGTGGAACAGCGGCGCGCTGACGAGCGGGGTCCACACCTTCCCGGCCGTGCTGCAGAGCAGCGGCCTCTACTCGCTGAGCAGCGTGGTCACCGTGCCCAGCAGCAGCCTGGGGACCCAGACGTACATCTGCAACGTGAACCACAAGCCCTCGAACACCAAGGTCGACAAGAAGGTGGAGCCCCCGAAGAGCTGCGACGGTACCCACACATGCCCACCGTGCCCAGGTACTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGTGAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA 94 15 FIG. 1AA sequence of HC of Ab A18EVQLLESGGGLVKPGQSLKLSCAASGETFTSYGMHWVRQPPGKGLEWVAVISYDGSYKYYADSVQGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSRLRSLLYFEWLSQGYFNPWGAGTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVtOYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYlCNVNHKPSNTKVDKKVEPPKSCDKTHTCPPCPGXELLGGPSVFLFPPKPKDTLMISRTPEVXCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGEPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK188 28 FIG. 1 AA sequence of LC of Ab A18EIVMTQSPDSLAVSLGERATINCKSSQSVTYNYKNYLAWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYRTPPTFGGGTKLDIKGSVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGE 145 n.a. see textAA sequence of LC CDR1 of Ab 044 QSITFDYKNYLA 146 n.a. see textAA sequence of LC CDR1 of F16 VK KSSQSVTFNYKNYLA 147 n.a. see textAA sequence of LC CDR2 of F16 VK WASARES 148 n.a. see textAA sequence of LC CDR3 of F16 VK QQHYRTPPT 68 n.a. see textAA sequence of HC CDR1 of Abs 044, SYAMH 069, 032, 031 69 n.a. see textAA sequence of HC CDR2 of Abs 044, VVSYDGNYKYYADVSQG 069, 032, 031 70n.a. see text AA sequence of HC CDR3 of Abs 044, DSRLRSLLYFEWLSQGYFNP069, 032, 031 71 n.a. see text AA sequence of LC CDR1 of Abs 032, 031QSITFNYKNYLA 72 n.a. see text AA sequence of LC CDR2 of Abs 044, 069,WGSYLES 032, 031 73 n.a. see textAA sequence of LC CDR3 of Abs 044, 069, QQHYRTPPS 032, 031 74 n.a.see text AA sequence of HC FR1 of Ab 069 QVQLLETGGGLVKPGQSLKLSCAASGFTFT75 n.a. see text AA sequence of HC FR2 of Ab 069 WVRQPPGKGLEWVA 76 n.a.see text AA sequence of HC FR3 of Ab 069RFTISRDNSKNTLYLQMNSLRAEDTAVYYCAK 77 n.a. see textAA sequence of HC FR4 of Ab 069 WGQGTTLTVSS 78 n.a. see textAA sequence of LC FR1 of Ab 069 DIQMTQSPSSLSASVGDRVTITCRSS 79 n.a.see text AA sequence of LC FR2 of Ab 069 WYQQKPGKAPKLLIY 80 n.a.see text AA sequence of LC FR3 of Ab 069GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC 81 n.a. see textAA sequence of LC FR4 of Ab 069 FGQGTKVEIK 82 n.a. see textAA sequence of HC FR1 of Ab 031 EVQLLESGGGLVKPGQSLKLSCAASGFTFT 83 n.a.see text AA sequence of LC CDR1 of Ab A18 et al. KSSQSVTYNYKNYLA 84 n.a.see text AA sequence of LC CDR2 of Ab A18 et al. WASTRES 85 n.a.see text AA sequence of LC CDR3 of Ab A18 et al. QQYYRTPPT 86 n.a.see text AA sequence of HC CDR1 of Ab A18 et al. SYGMH 87 n.a. see textAA sequence of HC CDR2 of Ab A18 et al. VISYDGSYKYYADSVQG 88 n.a.see text AA sequence of an HC CDR3 DSELRSLLYFEWLSQGYFNP 89 n.a. see textAA sequence of HC FR4 of Ab A18 et al. WGAGTTLTVSS 90 n.a. see textAA sequence of LC FR1 of Ab A18 et al. EIVMTQSPDSLAVSLGERATINC 91 n.a.see text AA sequence of LC FR2 of Ab A18 et al. WYQQKPGQPPKLLIY 92 n.a.see text AA sequence of LC FR3 of Ab A18 et al.GVPDRFSGSGSGTDFTLTISSLQAEDVAVYYC 93 n.a. see textAA sequence of LC FR4 of Ab A18 et al. FGGGTKLDIK 171 n.a. see textAA sequence of HC FR4 of Ab 078 et al. WGQGTTVTVSS 172 n.a. see textAA sequence of LC CDR1 of Ab 069 QSITFEYKNYLA 173 n.a. see textAA sequence of H3 HA1QDLPGNDNSTATLCLGHHAVPNGTLVKTITDDQIEVTNATELVQSSSTGKICNNPHRILDGIDCTLIDALLGDPHCDVFQNETWDLFVERSKAFSNCYPYDVPDYASLRSLVASSGTLEFITEGFTWTGVTQNGGSNACKRGPGSGFFSRLNWLTKSGSTYPVLNVTMPNNDNFDKLYIWG1HHPSTNQEQTSLYVQASGRVTVSTRRSQQTIIPNIGSRPWVRGLSSRISIYWTIVKPGDVLVINSNGNLIAPRGYFKMRTGKSSIMRSDAPIDTCISECITPNGSIPNDKPFQNVNKITYGACPKYVKQNTLKLATGMRNVPEKQTR 174 n.a. see textAA sequence of H3 HA2GLFGAIAGFIENGWEGMIDGWYGFRHQNSEGTGQAADLKSTQAAIDQINGKLNRVIEKTNEKFHQIEKEFSEVEGRIQDLEKYVEDTKIDLWSYNAELLVALENQHTIDLTDSEMNKLFEKTRRQLRENAEEMGNGCFKLYHKCDNACIESIRNGTYDHDVYRDEALN NRFQIKG 175n.a. FIG. 4 AA sequence of HC VR of FI6QVQLVQSGGGVVQPGRSLRLSCVASGFTFSTYAMHWVRQAPGRGLEWVAVISYDGNYKYYADSVKGFRSISRDNSNNTLHLEMNTLRTEDTALYYCAKDSQLRSLLYFEWLSQGYFDPWGQGTLVTVTS 176 n.a. FIG. 4 AA sequence of HC VR of FI370QVQLVQSGGGVVQPGRSLRLSCVASGFTFSTYAMHWVRQAPGRGLEWVAVISYDGNYKYYADSVKGFRSISRDNSNNTLHLEMNTLRTEDTALYYCAKDSQLRSLLYFEWLSQGYFDPWGQGTLVTVTS 177 n.a. FIG. 4 AA sequence of HC VR of FI6 variant 1QVQLVQSGGGVVQPGRSLRLSCVASGFTFSTYAMHWVRQAPGRGLEWVAVISYDGNYKYYADSVKGFRSISRDNSNNTLHLEMNTLRTEDTALYYCAKDSQLRSLLYFEWLSQGYFDPWGQGTLVTVTS 178 n.a. FIG. 4 AA sequence of HC VR of FI6 variant 3QVQLVQSGGGVVQPGRSLRLSCVASGFTFSTYAMHWVRQAPGRGLEWVAVISYDGNYKYYADSVKGFRSISRDNSNNTLHLEMNTLRTEDTALYYCAKDSQLRSLLYFEWLSQGYFDPWGQGTLVTVTS 179 n.a. FIG.4 AA sequence of HC VR of FI6/370QVQLVQSGGGVVQPGRSLRLSCVASGFTFSTYAMHWVRQAPGRGLEWVAVISYDGNYKYYADSVKGFRSISRDNSNNTLHLEMNTLRTEDTALYYCAKDSQLRSLLYFEWLSQGYFDPWGQGTLVTVTS 180 n.a. FIG. 4 AA sequence of kappa LC VR of FI6DIQMTSQPDSLAVSLGARATINCKSSQSVTFNYKNYLAWYQQKPGQPPKVLIYWASARESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQHYRTPPTFGQGTKVEIK 181 See textAA sequence of H1 HA1TNADTICIGYHANNSTDTVDTVLEKNVTVTHSVNLLEDSHNGKLCKLKGIAPLQLGKCNIAGWLLGNPECDLLLTASSWSYIVETSNSENGTCYPGDFIDYEELREQLSSVSSFEKFEIFPKTSSWPNHETTKGVTAACSYAGASSFYRNLLWLTKKGSSYPKLSKSYVNNKGKEVLVLWGVHHPPTGTDQQSLYQNADAYVSVGSSKYNRRFTPEIAARPKVRDQAGRMNYYWTLLEPGDTITFEATGNLIAPWYAFALNRGSGSGIITSDAPVHDCNTKCQTPHGAINSSLPFQNIHPVTIGECPKYVRSTKLRMATGLRNIPSIQS 182 See textAA sequence of H1 HA2GLFGAIAGFIEGGWTGMIDGWYGYHHQNEQGSGYAADQKSTQNAIDGITNKVNSVIEKMNTQFTAVGKEFNNLERRIENLNKKVDDGFLDIWTYNAELLVLLENERTLDFHDSNVRNLYEKVKSQLKNNAKEIGNGCFEFYHKCDDACMESVRNGTYDYPKYSEESKLNREEIDGVKLESMGVYQILAIYSTVASSLVLLVSLGAISFWMCSNGSLQCRICI

Examples Example 1. Designing of Anti-HA Antibodies

Human antibodies (IgG) targeting viral hemagglutinin (HA) werecomputationally designed. HA mediates viral binding to host cell surfacereceptor, and cell membrane fusion to the viral envelope, resulting inviral entry. The antibody molecules described herein were designed toblock HA's fusogenic activity.

All antibody constructs were based on human IgG1 structure (γ1 heavychain and κ light chain). Point mutations in the V_(H) (variable heavydomain) and V_(L) (variable light domain) were computationally designed.These mutations are located within or outside the CDRs (ComplementarityDetermining Regions). The mutations were designed, e.g., to modifyantigen binding properties (e.g., for stronger or weaker bindingaffinity), or to stabilize structure, or to improve expressionproperties, etc.

The heavy and light chain sequences of one antibody, called A18 isprovided in FIG. 1.

The heavy and light chain pairings for exemplary computationallydesigned antibodies are shown in Table 3, above in Detailed Description.

DNA sequences for the variable heavy chain and variable light chain foreach of antibodies Ab A18, Ab 031, Ab 032, Ab 044, Ab 014 and Ab 028 areprovided below.

VH16: (SEQ ID NO: 63) GAGGTACAGCTCCTCGAATCGGGAGGGGGACTGGTCAAACCCGGTCAATCGCTCAAACTCTCGTGTGCAGCGTCAGGTTTTACGTTCAGCTCATATGGGATGCACTGGGTCCGCCAGCCTCCGGGAAAGGGACTGGAGTGGGTGGCAGTCGTGTCGTATGACGGGAGCAATAAGTACTACGCCGATTCAGTGCAAGGTCGGTTTACCATTTCGAGGGATAACAGCAAGAACACGCTCTACTTGCAGATGAACTCACTTAGAGCGGAAGATACGGCTGTGTACTATTGCGCCAAAGACACAAAGCTGCGATCCCTGTTGTACTTCGAATGGTTGTCCTCGGGCTTGCTTGACTATTGGGGGCAGGGCGCCATGGTCACAGTATCCAGCGCGTCGACTAAGG GGCCC VL29:(SEQ ID NO: 64) GAGATCGTGATGACGCAGAGCCCCGATAGCCTCGCTGTCTCATTGGGGGAACGGGCCACGATTAACTGCAAATCCTCACAGTCGGTGACTTTCAGCTATAAGAATTACCTGGCATGGTATCAGCAGAAGCCGGGTCAACCCCCAAAACTGTTGATCTACTGGGCCTCCACACGCGAGTCGGGAGTCCCGGACCGATTTTCGGGTTCAGGGTCCGGCACTGACTTTACCCTCACAATTTCATCGCTTCAAGCGGAGGATGTAGCAGTGTACTATTGTCAGCAGTATTACAGAACACCTCCCACCTTCGGAGGGGGAACGAAACTTGACATCAAGGGATCC VL30: (SEQ ID NO: 65)GAGATCGTGATGACGCAGAGCCCCGATAGCCTCGCTGTCTCATTGGGGGAACGGGCCACGATTAACTGCAAATCCTCACAGTCGGTGACTTTCGACTATAAGAATTACCTGGCATGGTATCAGCAGAAGCCGGGTCAACCCCCAAAACTGTTGATCTACTGGGCCTCCACACGCGAGTCGGGAGTCCCGGACCGATTTTCGGGTTCAGGGTCCGGCACTGACTTTACCCTCACAATTTCATCGCTTCAAGCGGAGGATGTAGCAGTGTACTATTGTCAGCAGTATTACAGAACACCTCCCACCTTCGGAGGGGGAACGAAACTTGACATCAAGGGATCC VH15: (SEQ ID NO: 66)GAAGTGCAACTCCTCGAGTCAGGAGGAGGTTTGGTGAAACCGGGTCAGTCCTTGAAACTGAGCTGTGCAGCAAGCGGGTTCACGTTTACGTCGTACGGCATGCACTGGGTACGGCAGCCTCCCGGGAAGGGACTTGAATGGGTCGCCGTCATCTCATACGACGGGTCGTACAAATACTATGCGGATAGCGTGCAAGGTCGCTTCACAATTTCCCGGGACAATTCGAAGAATACACTGTATCTTCAGATGAACTCGCTCAGGGCTGAGGACACGGCGGTCTATTACTGCGCGAAGGATTCGCGACTCAGATCCCTTTTGTACTTTGAGTGGCTGTCGCAGGGGTATTTCAACCCATGGGGAGCCGGAACCACTTTGACCGTATCAAGCGCGTCAACAAAGG GGCCC VL28:(SEQ ID NO: 67) GAAATTGTAATGACGCAGAGCCCTGATAGCCTTGCCGTGTCCCTGGGTGAGAGGGCGACAATCAATTGTAAGTCATCACAGTCGGTCACGTACAACTACAAGAACTACCTGGCGTGGTATCAACAGAAACCCGGGCAGCCGCCCAAATTGCTCATCTATTGGGCTTCGACACGGGAGTCGGGTGTGCCAGACCGCTTCTCCGGGTCAGGATCGGGAACTGACTTCACGTTGACTATTTCGTCCCTCCAGGCAGAAGATGTAGCCGTCTACTATTGCCAACAGTATTACAGAACGCCGCCTACATTTGGAGGCGGGACCAAACTTGACATCAAGGGATCCGTGGCCGCCCCCAGCGTCTTCATCTTCCCGCCCAGCGACGAGCAGCTGAAGTCGGGCACGGCCAGCGTGGTGTGCCTCCTGAACAACTTCTACCCCCGCGAGGCGAAGGTCCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGGAACAGCCAGGAGAGCGTGACCGAGCAGGACTCGAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAGGCCGACTACGAGAAGCACAAGGTCTACGCCTGCGAGGTGACCCACCAGGGGCTCTCGAGCCCCGTGACCAAGAGCTTCAACCGGGGCGA GTGC VL52:(SEQ ID NO: 149) GACATTCAGATGACTCAGTCGCCTTCGTCATTGTCCGCCTCCGTGGGTGATAGGGTCACGATCACGTGCCGGAGCAGCCAGTCCATCACCTTCAATTACAAAAACTATTTGGCATGGTATCAACAGAAACCCGGAAAGGCGCCGAAGCTCCTGATCTACTGGGGTTCATATCTTGAGTCGGGGGTGCCGTCGAGATTTTCGGGCAGCGGATCAGGGACGGATTTCACGCTGACCATTTCGTCACTCCAGCCCGAGGACTTTGCGACATATTACTGTCAACAGCACTACAGGACACCCCCATCTTTCGGACAGGGGACTAAAGTAGAAATCAAGGGATCCGTGGCCGCCCCCAGCGTCTTCATCTTCCCGCCCAGCGACGAGCAGCTGAAGTCGGGCACGGCCAGCGTGGTGTGCCTCCTGAACAACTTCTACCCCCGCGAGGCGAAGGTCCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGGAACAGCCAGGAGAGCGTGACCGAGCAGGACTCGAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAGGCCGACTACGAGAAGCACAAGGTCTACGCCTGCGAGGTGACCCACCAGGGGCTCTCGAGCCCCGTGACCAAGAGCTTCAACCGGGGCGA GTGCTGA VL45:(SEQ ID NO: 150) GACATTCAGATGACTCAGTCGCCTTCGTCATTGTCCGCCTCCGTGGGTGATAGGGTCACGATCACGTGCCGGAGCAGCCAGTCCATCACCTTCAATTACAAAAACTATTTGGCATGGTATCAACAGAAACCCGGAAAGGCGCCGAAGCTCCTGATCTACTGGGGTTCATATCTTGAGTCGGGGGTGCCGTCGAGATTTTCGGGCAGCGGATCAGGGACGGATTTCACGCTGACCATTTCGTCACTCCAGCCCGAGGACTTTGCGACATATTACTGTCAACAGCACTACAGGACACCCCCATCTTTCGGACAGGGGACTAAAGTAGAAATCAAGGGATCCGTGGCCGCCCCCAGCGTCTTCATCTTCCCGCCCAGCGACGAGCAGCTGAAGTCGGGCACGGCCAGCGTGGTGTGCCTCCTGAACAACTTCTACCCCCGCGAGGCGAAGGTCCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGGAACAGCCAGGAGAGCGTGACCGAGCAGGACTCGAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAGGCCGACTACGAGAAGCACAAGGTCTACGCCTGCGAGGTGACCCACCAGGGGCTCTCGAGCCCCGTGACCAAGAGCTTCAACCGGGGCGA GTGCTGAGAATTC VH25:(SEQ ID NO: 151) CAGGTACAATTGCTTGAGACAGGTGGAGGACTCGTGAAGCCAGGTCAGTCATTGAAACTGAGCTGTGCCGCATCCGGGTTCACATTCACTTCCTACGCGATGCACTGGGTCCGCCAGCCTCCCGGAAAGGGACTTGAGTGGGTCGCTGTGGTATCGTATGATGGGAATTACAAATACTATGCAGACTCCGTGCAAGGCCGGTTTACGATTAGCAGGGACAACTCGAAGAATACCCTTTACCTCCAAATGAACTCGCTCCGAGCGGAGGACACGGCGGTGTATTACTGCGCGAAGGATTCACGGTTGAGATCGCTGCTCTATTTTGAATGGTTGTCACAGGGGTACTTCAACCCGTGGGGTCAGGGAACAACACTGACCGTCAGCTCAGCCTCGACTAAAGGGCCCAGCGTGTTCCCGCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGGACCGCCGCCCTGGGCTGCCTCGTCAAGGACTACTTCCCCGAGCCCGTGACCGTGTCGTGGAACAGCGGCGCGCTGACGAGCGGGGTCCACACCTTCCCGGCCGTGCTGCAGAGCAGCGGCCTCTACTCGCTGAGCAGCGTGGTCACCGTGCCCAGCAGCAGCCTGGGGACCCAGACGTACATCTGCAACGTGAACCACAAGCCCTCGAACACCAAGGTCGACAAGAAGGTGGAGCCCCCGAAGAGCTGCGACAAAACTCACACATGCCCACCGTGCCCAGGTACTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGTGAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA VH24: (SEQ ID NO: 152)GAAGTACAATTGCTTGAGTCGGGTGGAGGACTCGTGAAGCCAGGTCAGTCATTGAAACTGAGCTGTGCCGCATCCGGGTTCACATTCACTTCCTACGCGATGCACTGGGTCCGCCAGCCTCCCGGAAAGGGACTTGAGTGGGTCGCTGTGGTATCGTATGATGGGAATTACAAATACTATGCAGACTCCGTGCAAGGCCGGTTTACGATTAGCAGGGACAACTCGAAGAATACCCTTTACCTCCAAATGAACTCGCTCCGAGCGGAGGACACGGCGGTGTATTACTGCGCGAAGGATTCACGGTTGAGATCGCTGCTCTATTTTGAATGGTTGTCACAGGGGTACTTCAACCCGTGGGGTCAGGGAACAACACTGACCGTCAGCTCAGCCTCGACTAAAGGGCCCAGCGTGTTCCCGCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGGACCGCCGCCCTGGGCTGCCTCGTCAAGGACTACTTCCCCGAGCCCGTGACCGTGTCGTGGAACAGCGGCGCGCTGACGAGCGGGGTCCACACCTTCCCGGCCGTGCTGCAGAGCAGCGGCCTCTACTCGCTGAGCAGCGTGGTCACCGTGCCCAGCAGCAGCCTGGGGACCCAGACGTACATCTGCAACGTGAACCACAAGCCCTCGAACACCAAGGTCGACAAGAAGGTGGAGCCCCCGAAGAGCTGCGACGGTACCCACACATGCCCACCGTGCCCAGGTACTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGTGAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA

Each of the above sequences can be modified to include an ATCGATnucleotide sequence at the 5′ end, which will encode a variable heavychain or light chain polypeptide comprising Ile-Asp at the aminoterminus.

Example 2. Initial Formulation Study

This Example summarizes the initial formulation study results for anexemplary anti-HA antibody molecule described herein, e.g., Ab 044.

Procedure

Fourteen formulation matrices with different pH values with 40 mM sodiumphosphate-citrate buffer and with different compositions were prepared.The antibody sample was prepared by a sequence of Protein A affinitychromatography, anion exchange chromatography, and cation exchangechromatography. The purified samples were formulated in the appropriatesolutions using Ultrafiltration/Diafiltration. Aliquots of 4.0 mL ofantibody at 10 mg/mL in 30 mM Sodium Phosphate, 8.6 mM Citric Acid, 50m·M Histidine, 90 m·M NaCl, pH 6.0 were transferred into a 30K MWCOAmicon Ultra Centrifugal filter and centrifuged at 3600 RPM for 30minutes. Additional 2.0 mL of antibody at concentration of 10 mg/mL wasadded into each 30K MWCO Amicon Ultra Centrifugal filter and centrifugeat 3600 RPM for 40 minutes. The solution volume in each Amicon UltraCentrifugal filter was reduced to approximately 500 μL. Aliquots of 4.0ml of formulation matrix per filter were added (see Table 5 for theformulation components with 40 mM sodium phosphate/citrate buffer) andcentrifuged at 3600 RPM for 40 minutes. The solution volume in eachAmicon Ultra Centrifugal filter was reduced to approximately 500 μL.Additional 4.0 ml of formulation matrix per filter were added and thefilters were centrifuged at 3600 RPM for 45 minutes. The solution volumein each Amicon Ultra Centrifugal filter was reduced to approximately 400μL. After completion of the two buffer exchanges, it can be assumed thatto the final solution contained less than 1.5% of the original solutioncomposition. The final retentate volume was reduced from 6.0 mL toapproximately 400 μL, with a theoretical concentration of around 150mg/ml assuming no protein was lost to the membrane or precipitated out.

The retentate was then filtered through a 0.22 μM membrane. A280 and DSCwere conducted to measure protein concentrations and the conformationalstability of the antibody in each formulation, respectively. Each samplewas also divided into 4 portions in glass vials. The first 3 aliquotshave 65 μL solution. The remaining sample is in the 4th aliquot, withsample volume ranged between 65 and 350 μL. The first 3 aliquots werestored at 5° C., 45° C., and −70° C., respectively; the 4th portion wasstored at 5° C. The 1st three aliquots was pulled on day 7 and frozen at−70° C. and shipped on dry ice for analysis. These samples were analyzedwith size exclusion-high performance liquid chromatography (SEC-HPLC),and the results are summarized in Table 6. The Appearance test wasconducted for all of the 1st three aliquots on day 1 and day 7 exceptthe −70° C. samples on day 7 which was not thawed before shipping. Allof samples appeared clear without visible particles. The 4th aliquot wasstored at 5° C. for later analysis.

Results and Discussion

Table 5 shows formulation information, protein concentration,differential scanning calorimetry (DSC) peak temperatures, and the finalvolume after buffer exchange, while, Table 6 summarizes the sizeexclusion chromatography (SEC) results.

TABLE 5 Formulation information with A280 and DSC Data Front Main NaClTw80 Sucro Hist Arg Gly Peak Peak Volume Number pH mM % % % % % ° C. °C. mg/mL uL 1 5.5 150 0.05 2 0 0 2 Minor 77.9 75.0 385 2 6.0 50 0 2 2 02 Minor 78.1 90.0 410 3 6.0 150 0.05 0 0 0 0 No 77.3 106.0 284 4 7.0 500 0 0 0 0 No 76.3 72.0 262 5 5.5 50 0 0 2 2 0 67.5 75.5 78.0 510 6 7.0150 0.05 2 2 2 2 No 77.3 73.0 436 7 5.5 150 0 0 2 0 2 65.5 75.4 66.0 4868 6.5 150 0 0 2 2 2 No 77.3 80.0 382 9 6.0 50 0.05 0 0 0 0 No 76.8 47.0317 10 8.0 150 0.05 2 2 0 0 No 76.2 66.0 406 11 8.0 50 0 0 0 2 2 No 76.569.0 475 12 5.0 100 0.025 1 1 1 1 65.3 65.3 73.0 483 13 5.0 50 0.05 0 02 0 68.4 68.4 57.0 481 14 6.0 150 0.05 0 0 2 0 No 76.8 59.0 588 Sucro =Sucrose; His = Histindine; Arg = Arginine; Gly = Glycine

FIGS. 8A-8G show the DSC profile for all of the 14 formulation samples.Significant differences were observed between the formulations by DSC. Afront shoulder was clearly observed for formulation #5, 7, 12, and 13 atapproximately 70° C. for low pH samples (pH 5-5.5), indicating theanti-HA antibody molecule at lower pH denatured sooner as thetemperature increased.

Table 6 shows the summary of the overall protein recovery of theconcentration step and the Size Exclusion Chromatography (SEC) resultsof the 14 formulations stored at 3 temperatures. The recovery of theconcentration step (targeted 100 mg/ml) was calculated based on theamount of protein at the start and end of the process. The results ofthe SEC analytics are expressed as a main peak, containing the HAantibody monomer, as well as peaks containing High Molecular Weight(HMW) and Low Molecular Weight (LMW) species, consisting of aggregatesand breakdown products. Data show that all 42 samples tested consist toa very large proportion of monomeric species, as expressed by % MainPeak above 98%, when stored at 2-8° C. However, % High Molecular WeightSpecies (% HMWS), and/or % Low Molecular Species (% LMWS) increased morein some of the formulation upon storage at 45° C. for 1 week (#1, 3, 4,9, 10, 11, 12 and 14) in comparison with rest of the formulationbuffers. The results indicated that the levels of stress-inducedaggregation and degradation vary depending on the formulation buffer.

TABLE 6 Summary of % Recovery of Concentration Step and SEC Results # %Recovery Temp ° C. % HMWS % Main Peak % LMWS 1 48 −70 0.5 99.5 0 5 0.799.3 0 45 1.2 98.8 0 2 62 −70 0 100 0 5 0.1 99.9 0 45 0.2 99.8 0 3 50−70 0.1 99.7 0.1 5 0.1 99.9 0 45 0.3 98.7 1.0 4 32 −70 0.5 99.5 0 5 0.599.5 0 45 0.7 98.8 0.5 5 66 −70 0.2 99.8 5 0.2 99.8 45 Crystallized 6 53−70 0.6 99.4 0 5 0.6 99.4 0 45 0.9 99.0 0 7 54 −70 0.5 99.5 0 5 0.4 99.60 45 0.6 99.4 0 8 51 −70 0.5 99.6 0 5 0.5 99.5 0 45 0.7 99.3 0 9 25 −700.1 99.9 0.1 5 0.1 99.9 0.0 45 0.2 99.0 0.8 10 45 −70 0.7 99.1 0.2 5 0.799.1 0.2 45 1.2 97.7 1.2 11 55 −70 0.5 99.5 0 5 0.4 99.4 0.2 45 1.2 98.70.1 12 59 −70 0.5 99.4 0 5 0.6 99.4 0 45 1.3 98.7 0 13 46 −70 0.6 99.4 05 0.6 99.4 0 45 3.5 95.7 0.8 14 58 −70 0.5 99.5 0 5 0.5 99.5 0 45 0.998.4 0.7 % HMWS = % High Molecular Weight Species and % LMWS = % LowMolecular Species

This initial evaluation indicates that the antibody can be formulated upto 106 mg/ml and is stable within a wide range of pH and buffercompositions at 2-8° C. Differential Scanning calorimetry (DSC) datafrom unstressed samples and the SEC-HPLC data on stressed and unstressedsamples revealed differences between the formulations. Notably,Formulation#3-38.6 mM Sodium Phosphate-Citrate, 150 mM Sodium Chloride,pH 6.0, 0.05% Tween-80 samples reached a concentration of 106 mg/ml, didnot result in a front should by DSC and maintained >98% monomer uponheat stress by SEC-HPLC.

Example 3: Development of Stable Formulations for Antibody Drug Product

This Example summarizes the formulation development study for anexemplary anti-HA antibody molecule described herein, e.g., Ab 044, at25 mg/ml. A short-term thermal stressed stability study, a freeze/thawstudy, and an agitation study were performed to screen out the desiredformulation.

Five formulations were prepared at concentration of 25 mg/mL. Eachformulation was divided into several portions for different storageconditions, which include 4° C. and 45° C. for 2 weeks, freeze/thaw for1 and 3 cycles, and agitation for 16 hours at speed of 30 RPM by a cPCole-Parmer. Appearance, SEC, CE-SDS, A280, IEF and potency analyseswere conducted to evaluate the stability of these samples. Table 7 liststhe composition of the 5 formulations. It was found that the antibodymolecule was stable for up to 3 freeze/thaw cycles and overnightagitation. Among the 5 formulations, Formulation 1 and 4 were moststable. Formulation 1 was chosen as the final formulation for theantibody molecule based on the potential long term benefit of Tween-80.

TABLE 7 Composition of Formulations Formulation # Formulation Title #140 mM Citrate-Sodium Phosphate, 150 mM Sodium Chloride, pH 6.0, 0.025%Tween-80 #2 40 mM Citrate-Sodium Phosphate, 150 mM Sodium Chloride, pH6..5, 0.025% Tween-80 #3 40 mM Citrate-Sodium Phosphate, 1% Glycine, 75mM Sodium Chloride, pH 6.5, 0.025% Tween-80 #4 40 mM Citrate-SodiumPhosphate, 150 mM Sodium Chloride, pH 6.0. #5 40 mM Citrate-SodiumPhosphate, 75 mM Sodium Chloride, pH 6.5, 0.025% Tween-80

Summary of the Formulation Procedure

Citric Acid (JT Baker, Lot K42466) 100 mM, Sodium Phosphate DibasicHeptahydrate (Fisher, Lot 125720) 100 mM, and NaCl (JT Baker, LotL10472) 1.0 N were prepared. pH 6.0 buffer (100 mM) was prepared bymixing Citric Acid 100 mM and Sodium Phosphate Dibasic Heptahydrate 100mM at a proper ratio determined by a pH meter. pH 6.5 buffer (100 mM)was also prepared by mixing Citric Acid 100 mM and Sodium PhosphateDibasic Heptahydrate 100 mM at a proper ratio determined by the pHmeter. The 5 formulations were prepared without Tween-80 according toTable 8 and were QS to a final volume of 125 mL with water in agraduated cylinder.

TABLE 8 Recipe of Formulation Buffer without Tween-80 Formulation # 1 23 4 5 NaCl, 1N, mL; Final Conc = 75 or 18.8 18.8 9.4 18.8 9.4 150 nM pH6.0 100 mM, mL; Final Conc = 50 0 0 50 0 40 mM pH 6.5 100 mM, mL; FinalConc = 0 50 50 0 50 40 mM Glycine, g; Final Conc = 0% or 0 0 1.25 0 01.0%

After 125 mL of formulation buffers were prepared, the pH values werefurther adjusted by 5N NaOH or 5N HCl to pH 6.0 or 6.5. The antibodybulk drug substance (BDS) (4.5 mg/mL, 229.2 mL) was prepared by asequence of Protein A affinity chromatography, anion exchangechromatography, and cation exchange chromatography. The purified sampleswere formulated in the appropriate solutions usingUltrafiltration/Diafiltration.

Amicon Ultracel 30K Lot R2AA64948 (Max mL=15 mL) ultrafiltration tubeswere used to perform buffer exchange/concentration. Sample volumes wereadjusted according to A280 results with target value of 25 mg/mL. Table9 lists recoveries of the formulation samples. Formulation #1 and #4were combined.

TABLE 9 Recoveries of Formulation Total Concentration LoadingFormulation # Weight g mg/mL mg Recovery % #1 and #4 5.61 g 24.69 174.2679.5% #2 2.68 g 24.76 87.13 76.2% #3 2.71 g 24.82 87.13 77.2% #5 2.64 g24.97 87.13 75.7%

The combined formulation sample #1 and #4 was divided into equalportions as Formulation #1 and Formulation #4, respectively. DilutedTween-80 (JT Baker, Lot H35614) (3%) was spiked into the samples exceptFormulation #4 to reach a concentration of 0.025% Tween-80. Eachformulation sample was divided into multiple portions for testing ondifferent conditions of T=0, Agitation, 1 cycle of freeze/thaw, 3 cyclesof freeze/thaw, and for 2 weeks at 45° C.

Results

During the study, all samples were colorless without precipitation orvisible particles. Table 10 lists the A280 results. The proteinconcentration after storage at different conditions remained the same.

TABLE 10 A280 Results Sample ID mg/mL Sample ID mg/mL Sample ID mg/mLFormulation #1, T = 0 24.8 #1 T = 2 weeks 4° C. 24.5 #1 T = 2 weeks 45°C. 24.3 Formulation #1 1 F/T, T = 0 24.9 Formulation #1 3 F/T, T = 024.3 Formulation #1 Agitate, T = 0 24.4 Formulation #2, T = 0 24.6 #2 T= 2 weeks 4° C. 25.1 #2 T = 2 weeks 45° C. 24.3 Formulation #2 1 F/T, T= 0 24.9 Formulation #2 3 F/T, T = 0 25.0 Formulation #2 Agitate, T = 024.5 Formulation #3, T = 0 24.3 #3 T = 2 weeks 4° C. 24.9 #3 T = 2 weeks45° C. 24.3 Formulation #3 1 F/T, T = 0 24.7 Formulation #3 3 F/T, T = 024.5 Formulation #3 Agitate, T = 0 25.6 Formulation #4, T = 0 23.8 #4 T= 2 weeks 4° C. 24.5 #4 T = 2 weeks 45° C. 24.7 Formulation #4 1 F/T, T= 0 24.7 Formulation #4 3 F/T, T = 0 23.8 Formulation #4 Agitate, T = 024.9 Formulation #5, T = 0 24.8 #5 T = 2 weeks 4° C. 25.2 #5 T = 2 weeks45° C. 24.3 Formulation #5 1 F/T, T = 0 24.9 Formulation #5 3 F/T, T = 024.7 Formulation #5 Agitate, T = 0 24.6

Table 11 lists capillary electrophoresis-SDS (CE-SDS) results forreduced samples. The antibody molecule was stable over freeze/thaw andagitation based on the reduced CE-SDS results. The combined (heavy chainand light chain) Main peak % area after storage at different conditionsare listed. Formulations #1 and #4 showed improved stability as comparedto the other 3 formulations. FIG. 9 shows a representativeelectropherogram of CE-SDS for a reduced sample.

TABLE 11 Combined (HC and LC) Main Peak % of CE-SDS for Reduced SamplesFormulation # #1 #2 #3 #4 #5 CE Reduced HC + LC, 4 C., 2 wks 98.6 98.698.6 98.6 98.5 CE Reduced 96.9 96.4 95.7 97.2 96.4 Difference T = 2 wks4 C. - 1.7 2.2 2.9 1.4 2.1 T = 2 wks 45 C. CE Reduced HC + LC 4 C. 2 wks98.6 98.6 98.6 98.6 98.5 CE Reduced HC + LC 1 F./T 4 C. 98.7 98.6 98.798.4 98.6 2 wks CE Reduced HC + LC 3 F./T 4 C. 98.7 98.7 98.7 98.6 98.82 wks CE Reduced HC + LC Agitation 98.7 98.7 98.7 98.8 98.6 4 C. 2 wks

Table 12 lists CE-SDS results for non-reduced samples. The Main peak %area for IgG at different conditions are listed. The antibody moleculewas stable over freeze/thaw and agitation based on non-reduced CE-SDS.Formulations #1 and #4 appeared to maintain stability better than theother 3 formulations as demonstrated by the % Purity difference between4° C. and 45° C. storage for 2 weeks. FIG. 10 shows a representativeelectropherogram of CE-SDS for a non-reduced sample.

TABLE 12 Main Peak % of CE-SDS for Non-Reduced Samples Formulation # #1#2 #3 #4 #5 CE Non-Reduced IgG 4 C. 2 wks 97.0 97.3 97.4 97.1 97.1 CENon-Reduced IgG 45 C. 2 wks 92.0 91.0 90.6 91.7 91.4 Difference T = 2wks 4 C. - 5.0 6.3 6.8 5.4 5.7 T = 2 wks 45 C. CE Non-Reduced IgG 1 F./T4 C. 97.0 96.9 96.9 96.8 96.8 2 wks CE Non-Reduced IgG 3 F./T 4 C. 96.896.7 96.8 96.7 96.6 2 wks CE Non-Reduced IgG Agitation 96.6 96.4 96.696.4 96.4 4 C. 2 wks

Table 13 lists SEC-HPLC results as % peak area for monomer peak, highmolecular weight (HMW) peak and low molecular weight (LMW) peak fromsamples stored for 2 weeks at 4° C. and 45° C. The peak area %differences at 4° C. and 45° C. are also listed for differentformulations. Formulation #2 and #3 showed the most change upon stressat 45° C. FIG. 11 shows a representative SEC chromatogram.

TABLE 13 SEC Result for 2 Week Samples at 4° C. and 45° C. Sample ID HMW(%) Monomer (%) LMW (%) Formulation #1, T = 0 1.57 98.43 0 Formulation#1; 4 C. 2 wks 1.90 98.10 0 Formulation #1; 45 C. 2 wks 4.31 94.44 1.25Formulation #2, T = 0 1.81 98.19 0 Formulation #2; 4 C. 2 wks 1.97 98.030 Formulation #2; 45 C. 2 wks 4.70 94.03 1.27 Formulation #3, T = 0 1.5398.47 0 Formulation #3; 4 C. 2 wks 1.76 98.24 0 Formulation #3; 45 C. 2wks 4.70 94.21 1.09 Formulation #4, T = 0 1.68 98.32 0 Formulation #4; 4C. 2 wks 1.89 98.11 0 Formulation #4; 45 C. 2 wks 4.38 94.38 1.24Formulation #5, T = 0 1.64 98.36 0 Formulation #5; 4 C. 2 wks 1.96 98.040 Formulation #5; 45 C. 2 wks 4.29 94.54 1.17

Table 14 lists monomer % peak area from SEC for t=0 samples and afterfreeze/thaw (F/T) cycles or agitation Minimal changes were observed forthe different formulations after agitation, 1 cycle F/T, or 3 cyclesF/T.

TABLE 14 SEC Result for T = 0 Samples Name HMW (%) Monomer (%)Formulation #1; T = 0 1.57 98.43 Formulation #1; 1 F/T 1.64 98.36Formulation #1; 3 F/T 1.61 98.39 Formulation #1; Agitation 1.77 98.23Formulation #2; T = 0 1.81 98.19 Formulation #2; 1 F/T 1.94 98.06Formulation #2; 3 F/T 1.72 98.28 Formulation #2; Agitation 1.84 98.16Formulation #3; T = 0 1.53 98.47 Formulation #3; 1 F/T 1.55 98.45Formulation #3; 3 F/T 1.65 98.35 Formulation #3; Agitation 1.63 98.37Formulation #4; T = 0 1.68 98.32 Formulation #4; 1 F/T 1.53 98.47Formulation #4; 3 F/T 1.68 98.32 Formulation #4; Agitation 1.70 98.30Formulation #5; T = 0 1.64 98.36 Formulation #5; 1 F/T 1.65 98.35Formulation #5; 3 F/T 1.79 98.21 Formulation #5; Agitation 1.65 98.35

Isoelectric focusing (IEF) was conducted to analyze all samples. A majorband at the isoelectric point (pI) around 9.0 was observed for all ofthe samples. More degradation was observed for 45° C. samples (moreacidic bands noted by eyes). No significant difference was observedamong all the formulations. FIG. 12 shows a representative IEF gelimage.

Based on the results of non-reduced CE-SDS and SEC, t=O and t=2 wks 45°C. samples in formulation #3 were chosen to measure potency byHemagglutinin (HA) binding ELISA since the antibody molecule in thisformulation buffer had the most changes after storage at 45° C. for 2weeks. The results are summarized in Table 15. The data indicated nosignificant changes in potency (% Activity) between t=0 and the stressedsample considering the assay precision. Therefore, it is reasonable todraw the similar conclusion of no change in the antibody moleculepotency upon stress at 45° C. for 2 weeks in all of the formulationstested.

TABLE 15 Potency Result for t = 0 and 2 wks 45° C. Samples inFormulation #3 Sample % Activity T = 0 123 T = 2 wks 45 C. 97 Average %Activity 110 Standard Deviation (%) 18.4 % RSD 16.7

The results indicate that the antibody molecule was stable up to 3freeze/thaw cycles and overnight agitation. Among the 5 formulations,Formulation 1 and 4 were most stable. Formulation 1 was chosen as thefinal formulation for the antibody molecule based on the potential longterm benefit of Tween-80. The formulation contains 40 mM Citrate-SodiumPhosphate, 150 mM sodium chloride, 0.025% polysorbate-80, pH 6.0.

Example 4: Population Pharmacokinetic and Viral Dynamic Modeling ofVIS410 in a Human Challenge Model

A population pharmacokinetic (popPK) and influenza viral dynamic modelwere developed to support the VIS410 clinical program (e.g., using aformulation described herein), integrating data from a Phase 1 healthyvolunteer and a Phase 2a human influenza challenge study. VIS410 is alsoknown as Ab 044 herein.

Methods

Nasal and serum PK data from a Phase 1 study (N=30, single IV doses 2-50mg/kg) and a Phase 2a study (N=33, single IV doses of 2300 and 4600 mg)were used to develop the popPK model. In the Phase 2a study, volunteerswere inoculated intranasally with an attenuated influenza A (H1N1)strain, and received placebo or VIS410 24 h post-inoculation. Frequentnasal viral load (qPCR and TCID₅₀), serum and nasal PK were measured.The pharmacodynamic analysis included viral load data fromintent-to-treat infected subjects (ITT): placebo (n=7), 2300 mg (n=22),4600 mg (n=4). All analyses were performed in NONMEM 7.3 and qPCR andTCID50₅₀ were modeled separately; BLQ data were handled using the M3method, with predictive performance evaluated using NPDE (in R).

Results

A 3-compartment model adequately described PK with first-orderdistribution of VIS410 between nasal and central compartments (mean (%RSE) CL_(D) serum-to-nasal 0.04 (19.5%) mL/h; and nasal-to-serum 1.95(17.1%) mL/h). Body weight was the only covariate that was retained inthe popPK model. Other covariates tested included gender, age andinfection status, but were non-influential. A 92% reduction in viralload AUC by qPCR was observed at the 2300 mg dose compared to placebo(p<0.05). Viral dynamics in placebo and ITT subjects were wellcharacterized by a modified viral dynamic model comprising virus, targetepithelial cells, non-productive and productive infected cells; mAb drugeffect was modeled as inhibiting membrane fusion in the nasalcompartment, via an E_(max) function (mean (% RSE) EC₅₀ qPCR=1.96 (13)μg/mL and EC₅₀ TCID₅₀=18.4 (2.6) μg/mL).

In summary, VIS410 demonstrated PK generally typical of IgG1 mAbs, andpotent antiviral activity compared to placebo in the H1N1 humanchallenge model. A semi-mechanistic popPK model, which links mAb nasalconcentrations to influenza viral dynamics based on the VIS410 mechanismof action was successfully developed. The model describes serum andnasal PK, with impact on viral load, and was used to support doseselection for future clinical development across a spectrum ofpopulations. This approach may be extended to other mAbs targetedagainst influenza viral infections.

Additional examples are disclosed in International ApplicationPublication No. WO2013/170139, U.S. Pat. No. 8,877,200, U.S. Pat. No.9,096,657, and U.S. Patent Application Publication No. US 2013/0302349.The contents of the aforesaid publications are incorporated by referencein their entirety.

INCORPORATION BY REFERENCE

All publications, patents, and patent applications mentioned herein arehereby incorporated by reference in their entirety as if each individualpublication, patent or patent application was specifically andindividually indicated to be incorporated by reference. In case ofconflict, the present application, including any definitions herein,will control.

EQUIVALENTS

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, many equivalents to the specificembodiments of the invention described herein. Such equivalents areintended to be encompassed by the following claims.

What is claimed is:
 1. A formulation comprising 20 mg/mL to 60 mg/mL ofan anti-HA antibody molecule, 20 mM to 60 mM citrate-sodium phosphate,and 50 mM to 200 mM sodium chloride, wherein the formulation has a pH of6 to 6.5, and wherein the antibody molecule comprises: (a) a heavy chain(HC) immunoglobulin variable region segment comprising: an HC CDR1comprising the sequence S-Y-A-M-H (SEQ ID NO: 68); an HC CDR2 comprisingthe sequence V-V-S-Y-D-G-N-Y-K-Y-Y-A-D-S-V-Q-G (SEQ ID NO: 69); and anHC CDR3 comprising the sequence D-S-R-L-R-S-L-L-Y-F-E-W-L-S-Q-G-Y-F-N-P(SEQ ID NO: 70); and (b) a light chain (LC) immunoglobulin variableregion segment comprising: an LC CDR1 comprising the sequenceQ-S-I-T-F-D-Y-K-N-Y-L-A (SEQ ID NO: 145); an LC CDR2 comprising thesequence W-G-S-Y-L-E-S (SEQ ID NO: 72); and an LC CDR3 comprising thesequence Q-Q-H-Y-R-T-P-P-S (SEQ ID NO: 73).
 2. The formulation of claim1, further comprising 0.01% to 0.05% polysorbate 80,
 3. The formulationof claim 1, comprising 25 mg/mL to 50 mg/mL of the antibody molecule. 4.The formulation of claim 1, comprising 25 mg/mL of the antibodymolecule.
 5. The formulation of claim 1, wherein the antibody moleculecomprises a heavy chain immunoglobulin variable region segmentcomprising SEQ ID NO: 25, or an amino acid sequence that differs by nomore than 1, 2, 3, 4, or 5 amino acids therefrom.
 6. The formulation ofclaim 1, wherein the antibody molecule comprises a light chainimmunoglobulin variable region segment comprising SEQ ID NO: 52, or anamino acid sequence that differs by no more than 1, 2, 3, 4, or 5 aminoacids therefrom.
 7. The formulation of claim 1, wherein the antibodymolecule comprises: (a) a heavy chain immunoglobulin variable regionsegment comprising SEQ ID NO: 25, or an amino acid sequence that differsby no more than 1, 2, 3, 4, or 5 amino acids therefrom; and (b) a lightchain immunoglobulin variable region segment comprising SEQ ID NO: 52,or an amino acid sequence that differs by no more than 1, 2, 3, 4, or 5amino acids therefrom.
 8. The formulation of claim 1, wherein theantibody molecule comprises a heavy chain immunoglobulin variable regionsegment comprising SEQ ID NO: 25, a light chain immunoglobulin variableregion segment comprising SEQ ID NO: 52, or both.
 9. The formulation ofclaim 1, comprising 30 mM to 50 mM citrate-sodium phosphate.
 10. Theformulation of claim 1, comprising 40 mM citrate-sodium phosphate. 11.The formulation of claim 1, comprising 75 mM to 150 mM sodium chloride.12. The formulation of claim 1, comprising 75 mM sodium chloride. 13.The formulation of claim 1, comprising 150 mM sodium chloride.
 14. Theformulation of claim 1, which has a pH of
 6. 15. The formulation ofclaim 1, which has a pH of 6.5.
 16. The formulation of claim 1, furthercomprising 0.025% polysorbate
 80. 17. The formulation of claim 1,further comprising 0.5% to 2% glycine.
 18. The formulation of claim 1,further comprising 1% glycine.
 19. The formulation of claim 1,comprising 25 mg/mL of the antibody molecule, 40 mM citrate-sodiumphosphate, 150 mM sodium chloride, 0.025% polysorbate 80, wherein theformulation has a pH of
 6. 20. The formulation of claim 1, comprising 25mg/mL of the antibody molecule, 40 mM citrate-sodium phosphate, 150 mMsodium chloride, 0.025% polysorbate 80, wherein the formulation has a pHof 6.5.
 21. The formulation of claim 1, comprising 25 mg/mL of theantibody molecule, 40 mM citrate-sodium phosphate, 150 mM sodiumchloride, 0.025% polysorbate 80, 1% glycine, wherein the formulation hasa pH of
 6. 22. The formulation of claim 1, comprising 25 mg/mL of theantibody molecule, 40 mM citrate-sodium phosphate, 150 mM sodiumchloride, wherein the formulation has a pH of
 6. 23. The formulation ofclaim 1, comprising 25 mg/mL of the antibody molecule, 40 mMcitrate-sodium phosphate, 75 mM sodium chloride, 0.025% polysorbate 80,wherein the formulation has a pH of 6.5.
 24. The formulation of claim 1,which is a liquid formulation.
 25. A container comprising theformulation of claim
 1. 26. The container of claim 25, comprising 10 mLto 50 mL of the formulation.
 27. The container of claim 25, comprising20 mL to 40 mL of the formulation.
 28. The container of claim 25, whichis a vial.
 29. A method of preparing a composition for administration toa subject, the method comprises combining the formulation of claim 1with a solution suitable for intravenous administration.
 30. The methodof claim 29, wherein the solution comprises saline.
 31. The method ofclaim 30, wherein the solution further comprises dextrose.
 32. Themethod of claim 29, wherein 2000 mg to 5000 mg of the antibody moleculeis combined with the solution.
 33. The method of claim 29, wherein 2300mg to 4600 mg of the antibody molecule is combined with the solution.34. The method of claim 29, wherein the formulation is combined with thesolution in an intravenous solution bag.
 35. A method of treating orpreventing influenza, the method comprising administering to a subjecthaving influenza, or at risk of having influenza, an effective amount ofthe formulation of claim 1, thereby treating or preventing influenza.36. The method of claim 35, wherein the formulation is administered tothe subject intravenously.
 37. A container comprising 200 mL to 300 mLof a solution comprising an anti-HA antibody molecule, wherein thesolution is suitable for intravenous administration, wherein theantibody molecule is present at a concentration of 5 mg/mL to 20 mg/mL,and wherein the antibody molecule comprises: (a) a heavy chain (HC)immunoglobulin variable region segment comprising: an HC CDR1 comprisingthe sequence S-Y-A-M-H (SEQ ID NO: 68); an HC CDR2 comprising thesequence V-V-S-Y-D-G-N-Y-K-Y-Y-A-D-S-V-Q-G (SEQ ID NO: 69); and an HCCDR3 comprising the sequence D-S-R-L-R-S-L-L-Y-F-E-W-L-S-Q-G-Y-F-N-P(SEQ ID NO: 70); and (b) a light chain (LC) immunoglobulin variableregion segment comprising: an LC CDR1 comprising the sequenceQ-S-I-T-F-D-Y-K-N-Y-L-A (SEQ ID NO: 145); an LC CDR2 comprising thesequence W-G-S-Y-L-E-S (SEQ ID NO: 72); and an LC CDR3 comprising thesequence Q-Q-H-Y-R-T-P-P-S (SEQ ID NO: 73).
 38. The container of claim37, comprising 250 mL of a solution comprising the antibody molecule.39. The container of claim 37, wherein the antibody molecule is presentat a concentration of 8 mg/mL to 16 mg/mL.
 40. The container of claim37, wherein the antibody molecule comprises a heavy chain immunoglobulinvariable region segment comprising SEQ ID NO: 25, a light chainimmunoglobulin variable region segment comprising SEQ ID NO: 52, orboth.
 41. The container of claim 37, which is an intravenous (IV)solution bag.